山羊角膜上皮细胞与角膜内皮细胞分离培养研究
|
摘要
各种角膜疾病和外科损伤都会导致角膜细胞受损,当损伤超过其生理代偿时,就会发生角膜病变,严重时会导致角膜盲。目前角膜移植是治疗此类角膜疾病比较成熟的方法,但受到了供体角膜缺乏、术后并发症、角膜供体老龄化等条件的限制。将自体或异体角膜细胞体外培养构建组织工程化人工角膜移植,既可解决临床应用供体短缺问题,又可为角膜体外药理或生理研究提供大量材料。本研究通过体外分离培养山羊角膜上皮细胞和角膜内皮细胞,并研究其体外生长特性和接种羊膜载体培养情况,为组织工程人工角膜的构建奠定理论基础。
1.山羊角膜上皮细胞的体外分离培养与鉴定
通过比较组织块和酶消化法培养山羊原代角膜缘上皮细胞,发现酶消化法更适合原代角膜缘上皮细胞的分离纯化培养。获取的山羊角膜缘上皮细胞形态为多角形,具有较强的增殖能力,体外培养能传20多代,细胞冻存后仍具有细胞冻存前的形态和活力。经免疫组化和RT-PCR检测,培养的角膜缘上皮细胞表达K3/K12、p63、Cx43、ABCG2和PCNA,具有角膜上皮细胞和角膜缘干细胞表型,说明分离培养的细胞为角膜缘上皮细胞和角膜缘干细胞混合物,可为组织工程化人工角膜上皮的构建提供种子细胞。
2.组织工程化人工角膜上皮的构建
在去上皮细胞人羊膜上接种关中奶山羊角膜缘组织块或培养的关中奶山羊角膜缘上皮细胞构建的组织工程角膜上皮,并对其进行组织形态学、超微结构观察和免疫组化检测。结果显示,角膜缘组织块和角膜上皮细胞在羊膜上培养15 d均可形成6～7层结构,细胞内含丰富的细胞器和糖原颗粒,细胞间有大量的桥粒连接,细胞与羊膜间形成半桥粒连接,构建的复合角膜上皮p63染色阳性。比较角膜缘组织块在去羊膜上皮细胞的新鲜羊膜、冻存30 d羊膜、冻存60 d羊膜和冻存180 d羊膜上生长情况发现,从角膜缘组织块脱出的细胞生长面积在冻存30 d羊膜上最大,而在新鲜羊膜上细胞生长面积最小。比较角膜缘组织块在冻存60 d去上皮羊膜和未去上皮羊膜上生长情况发现,从角膜缘组织块脱出的细胞生长面积在去上皮羊膜上显著大于未去上皮羊膜上。说明用组织块法和种传代细胞法均能构建组织工程化人工角膜上皮,细胞培养载体需用冻存30 d左右去上皮细胞的羊膜,为组织工程化人工角膜上皮的构建研究奠定理论基础。
3.山羊角膜内皮细胞的体外分离培养与鉴定
比较组织块法、胰酶消化法、dispase酶消化法和dispase酶+胰酶消化法培养山羊原代角膜内皮细胞。dispase酶+胰酶消化法为角膜内皮细胞原代分离纯化的最适方法,分离的细胞在体外能连续传20多代,细胞冻存后解冻培养能保持冻存前细胞形态和活力。培养的细胞经扫描电镜和透射电镜观察,具有正常角膜内皮细胞特征,免疫组化染色和RT-PCR检测角膜内皮细胞表达神经元标志NSE和β3-tublin、星形胶质细胞标志GFAP、细胞间连接蛋白ZO-1和Cx43,不表达神经干细胞标志物Nestin、角膜缘干细胞标志p63和角膜上皮细胞标志K3/K12。比较细胞培养板包被物对角膜内皮细胞贴壁和增殖能力影响发现,Ⅳ胶原、多聚赖氨酸和明胶均能明显促进细胞贴壁和增殖,Ⅳ胶原和明胶组强于多聚赖氨酸组,而Ⅳ胶原组和明胶组之间差异不显著,提示可选择价格便宜的明胶作为细胞培养板用包被物。角膜内皮细胞培养液筛选发现:与NBS相比,FBS能更好的促进角膜内皮细胞增殖和维持其形态;培养液中添加bFGF能显著促进细胞增殖,但能促进细胞形态向成纤维形转变;添加EGF对细胞增殖影响不大,但能促进细胞形态改变;添加硫酸软骨素显著抑制细胞增殖,但能较好的维持细胞形态;添加Vc对细胞增殖和形态均无明显影响。将传代培养的角膜内皮细胞种羊膜上培养,经组织切片、透射和扫描电镜观察发现细胞在羊膜上具有形成单层的能力。说明分离培养的角膜内皮细胞具有正常角膜内皮细胞特征,能体外传代培养,在羊膜载体上具有形成单层的能力,可为构建组织工程化人工角膜内皮和角膜内皮细胞的体外实验研究提供种子细胞。
4.山羊角膜内皮细胞人端粒酶逆转录酶基因转染
将人端粒酶逆转录酶基因(hTERT)转入培养至7代的角膜内皮细胞内,通过免疫组化和RT-PCR检测证明转染的hTERT在山羊角膜内皮细胞内表达。通过传代培养观察,转染hTERT的角膜内皮细胞体外能传20多代,与未转染hTERT的角膜内皮细胞相比,并未显著提高其增殖能力。说明将人端粒酶基因转入山羊角膜内皮细胞内,未能明显延长细胞寿命。
All kinds of cornea diseases and traumas can result in corneal cells damaging. When the number of damaged-cells exceeds the threshold of corneal physiological compensation, pathologic changes will occur on the cornea and the sufferer can lose his sight eventually. Although corneal transplantation is regarded as an effective method curing the corneal diseases which induced the number of corneal cells by serious decreasing, the technology is not applied fully by some problems, such as the shortage of donors’cornea, the aging of donors and the syndrome after transplantation. Tissue engineering cornea which is regenerated with cultured cells from autograft or allograft in vitro can overcome the above-mentioned problems and is considered as the substitute of the cornea of dornors. It can be applied to the treatment of cornea diseases and the pharmacological or physiological research about cornea in vitro. To provide references for the rebuilding of tissue engineering artificial cornea, goat corneal epithelial cells and endothelial cells were isolated and cultured in vitro, the growth characteristic in culture dishes and on amniotic membrane were evaluated. We obtained the following results after carrying on the careful experiments.
1. Isolation, culture and identification of goat corneal epithelial cells in vitro
In this experiment, the explant culture and enzyme digestion method that used to isolated goat corneal epithelial cells separately were assessed. The result shows the enzyme digestion method was more suitable for isolation and culture of limbal epithelial cells than explant culture. Isolated Limbal epithelial cells gained by enzyme digestion method appear polygonal shapes and better proliferation and these cells can proliferate for 20 passages in vitro. Significant difference of morphology and proliferating capacity between pre-cryopreserved corneal epithelial cells and the ones after resuscitation was not found. The results of Immunostaining and RT-PCR showed that the cultured cells expressed some phenotype-characteristics of corneal epithelial cells and limbal stem cell, for instance, K3/K12, p63, Cx43, ABCG2 and PCNA. Therefore these isolated cells were made up of limbal epithelial cells and limbal stem cells and could be employed to rebuild tissue engineering corneal epithelium.
2. Reconstruction of tissue engineering corneal epithelium
Limbal explants and cultured limbal epithelial cells of Guanzhong dairy goats were seeded on the human denuded amniotic membrane for reconstructing tissue engineering corneal epithelium, whose histological morphology, ultrastructure and immunocytochemiacal characteristic were observed. Results showed that limbal stem cell could proliferate on amniotic membrane and form 6～7 layers of stratified epithelium. The cells of stratified epithelium contained abundant organelles and glycongenosomes, and there were a lot of desmosomes and hemidemosome between cells or cells and amnioticmembrane. Immunostaining of paraffinic slice for p63 was positive. Comparing the growth performance of limbal epithelial cells from limbal explants seeded respectively on uncryo-preserved, cryo-preserved 30-day, cryo-preserved 90-day, cryo-preserved 180-day human amniotic membrane which had been removed of epithelium, we concluded that the cellular outgrowing area from limbal explants seeded on cryo-preserved 30-day human amniotic membrane was biggest, the cellular outgrowing area from limbal explants seeded on fresh human amniotic membrane was smallest. The growth performance of limbal epithelial cells seeded on cryo-preserved 60-day human amniotic membrane removed of epithelium and unremoved of epithelium was compared, the results showed that the cellular outgrowing area from limbal explants seeded on denuded amniotic membrane were significantly larger than that of intact amniotic membrane and implicated that both limbal explants and cultured limbal epithelial cells seeded on denuded human amniotic membrane can be used to reconstruct tissue engineering epithelium, the 30-day denuded amniotic membrane were most suitable to growth of limbal epithelial cells. The results provided evidences for studying tissue engineering artificial cornea.
3. Isolation, Culture and Identification of Goat Corneal Endothelial Cells
Four methods including explant culture, trypsin digestion, dispase digestion and dispase with trypsin digestion method were employed to isolate goat corneal endothelial cells (GCECs)in this experiment. The effectiveness of the methods to isolate GCECs was assessed. It was an optimum method to isolate GCECs in vitro using dispase with trypsin digestion. The obtained cells by the method could be cultured for about 20-passage. The resuscitated cryo-preserved GCECs performed the same morphology and proliferating capacity as that of cryo-preserved GCECs. Resorting to scan and transmission electron microscope, we saw that the morphology of isolated GCECs resembled that of GCECs in vivo. The cultured GCECs in vitro were measured by immunocytochemical staining and RT-PCR, of which results showed that the cells expressed NSE andβ3-tublin of neuronal marker, GFAP of astroglial cell marker, ZO-1 and Cx43 of cell connecting protein marker,but not expressed Nestin of neuronal marker, p63 of limbal stem cell marker, K3/K12 of corneal epithelial cell marker. The influence of different substances coating plastic ware,Ⅳcollagen, gelatin and poly-L-lysine, on the adherence and proliferation of GCECs was assessed in these experiments. It was found thatⅣcollagen , gelatin and poly-L-lysine greatly improved the adherence and proliferation of GCECs.Ⅳcollagen and gelatin could promote GCECs adherence and proliferation more remarkably than poly-L-lysine, but there was no distinct difference betweenⅣcollagen and gelatin. While gelatin is less expensive thanⅣcollagen and more appropriate for GCECs culture in vitro. Basing on the experiment about optimizing culture medium for culturing GCECs, we have obtained following conclusions. Both FBS and NBS can promote cell proliferation and maintain cellular morphology, but FBS was more appropriate. Although bFGF could also promote cell proliferation remarkably, it could also change the morphology of GCECs into fibroblast-like. EGF could not promote cell proliferation, while also influenced cellular morphology. Chondroitin sulfate could inhibit cell proliferation and maintain cellular morphology. Vitamin C could not influence cell proliferation and cellular morphology. By optical microscope, scan and transmission electron microscope, we could conclude that cultured GCECs in vitro were seeded on the human denuded amniotic membrane could form a monoptychial tissue on amniotic membrane. The result also implicated the isolated GCECs had the characteristics of native GCECs and could be cultured in vitro to form a monoptychial on amniotic membrane. The cells can be used for reconstrcting tissue engineering epithelium.
4. Transfection of hTERT gene in GCECs
The human telomerase reverse transcriptase(hTERT) gene was transferred into GCECs. The results of immunocytochemical staining and RT-PCR showed that hTERT gene was expressed in GCECs, which could proliferate for 20 passages after being transfected by hTERT gene. But there was no significant difference of cellular proliferation between transfected cells and untransfected cells. The study implicated that the proliferation of GCECs could not be promoted by transfecting hTERT gene into GCECs.
1.山羊角膜上皮细胞的体外分离培养与鉴定
通过比较组织块和酶消化法培养山羊原代角膜缘上皮细胞,发现酶消化法更适合原代角膜缘上皮细胞的分离纯化培养。获取的山羊角膜缘上皮细胞形态为多角形,具有较强的增殖能力,体外培养能传20多代,细胞冻存后仍具有细胞冻存前的形态和活力。经免疫组化和RT-PCR检测,培养的角膜缘上皮细胞表达K3/K12、p63、Cx43、ABCG2和PCNA,具有角膜上皮细胞和角膜缘干细胞表型,说明分离培养的细胞为角膜缘上皮细胞和角膜缘干细胞混合物,可为组织工程化人工角膜上皮的构建提供种子细胞。
2.组织工程化人工角膜上皮的构建
在去上皮细胞人羊膜上接种关中奶山羊角膜缘组织块或培养的关中奶山羊角膜缘上皮细胞构建的组织工程角膜上皮,并对其进行组织形态学、超微结构观察和免疫组化检测。结果显示,角膜缘组织块和角膜上皮细胞在羊膜上培养15 d均可形成6～7层结构,细胞内含丰富的细胞器和糖原颗粒,细胞间有大量的桥粒连接,细胞与羊膜间形成半桥粒连接,构建的复合角膜上皮p63染色阳性。比较角膜缘组织块在去羊膜上皮细胞的新鲜羊膜、冻存30 d羊膜、冻存60 d羊膜和冻存180 d羊膜上生长情况发现,从角膜缘组织块脱出的细胞生长面积在冻存30 d羊膜上最大,而在新鲜羊膜上细胞生长面积最小。比较角膜缘组织块在冻存60 d去上皮羊膜和未去上皮羊膜上生长情况发现,从角膜缘组织块脱出的细胞生长面积在去上皮羊膜上显著大于未去上皮羊膜上。说明用组织块法和种传代细胞法均能构建组织工程化人工角膜上皮,细胞培养载体需用冻存30 d左右去上皮细胞的羊膜,为组织工程化人工角膜上皮的构建研究奠定理论基础。
3.山羊角膜内皮细胞的体外分离培养与鉴定
比较组织块法、胰酶消化法、dispase酶消化法和dispase酶+胰酶消化法培养山羊原代角膜内皮细胞。dispase酶+胰酶消化法为角膜内皮细胞原代分离纯化的最适方法,分离的细胞在体外能连续传20多代,细胞冻存后解冻培养能保持冻存前细胞形态和活力。培养的细胞经扫描电镜和透射电镜观察,具有正常角膜内皮细胞特征,免疫组化染色和RT-PCR检测角膜内皮细胞表达神经元标志NSE和β3-tublin、星形胶质细胞标志GFAP、细胞间连接蛋白ZO-1和Cx43,不表达神经干细胞标志物Nestin、角膜缘干细胞标志p63和角膜上皮细胞标志K3/K12。比较细胞培养板包被物对角膜内皮细胞贴壁和增殖能力影响发现,Ⅳ胶原、多聚赖氨酸和明胶均能明显促进细胞贴壁和增殖,Ⅳ胶原和明胶组强于多聚赖氨酸组,而Ⅳ胶原组和明胶组之间差异不显著,提示可选择价格便宜的明胶作为细胞培养板用包被物。角膜内皮细胞培养液筛选发现:与NBS相比,FBS能更好的促进角膜内皮细胞增殖和维持其形态;培养液中添加bFGF能显著促进细胞增殖,但能促进细胞形态向成纤维形转变;添加EGF对细胞增殖影响不大,但能促进细胞形态改变;添加硫酸软骨素显著抑制细胞增殖,但能较好的维持细胞形态;添加Vc对细胞增殖和形态均无明显影响。将传代培养的角膜内皮细胞种羊膜上培养,经组织切片、透射和扫描电镜观察发现细胞在羊膜上具有形成单层的能力。说明分离培养的角膜内皮细胞具有正常角膜内皮细胞特征,能体外传代培养,在羊膜载体上具有形成单层的能力,可为构建组织工程化人工角膜内皮和角膜内皮细胞的体外实验研究提供种子细胞。
4.山羊角膜内皮细胞人端粒酶逆转录酶基因转染
将人端粒酶逆转录酶基因(hTERT)转入培养至7代的角膜内皮细胞内,通过免疫组化和RT-PCR检测证明转染的hTERT在山羊角膜内皮细胞内表达。通过传代培养观察,转染hTERT的角膜内皮细胞体外能传20多代,与未转染hTERT的角膜内皮细胞相比,并未显著提高其增殖能力。说明将人端粒酶基因转入山羊角膜内皮细胞内,未能明显延长细胞寿命。
All kinds of cornea diseases and traumas can result in corneal cells damaging. When the number of damaged-cells exceeds the threshold of corneal physiological compensation, pathologic changes will occur on the cornea and the sufferer can lose his sight eventually. Although corneal transplantation is regarded as an effective method curing the corneal diseases which induced the number of corneal cells by serious decreasing, the technology is not applied fully by some problems, such as the shortage of donors’cornea, the aging of donors and the syndrome after transplantation. Tissue engineering cornea which is regenerated with cultured cells from autograft or allograft in vitro can overcome the above-mentioned problems and is considered as the substitute of the cornea of dornors. It can be applied to the treatment of cornea diseases and the pharmacological or physiological research about cornea in vitro. To provide references for the rebuilding of tissue engineering artificial cornea, goat corneal epithelial cells and endothelial cells were isolated and cultured in vitro, the growth characteristic in culture dishes and on amniotic membrane were evaluated. We obtained the following results after carrying on the careful experiments.
1. Isolation, culture and identification of goat corneal epithelial cells in vitro
In this experiment, the explant culture and enzyme digestion method that used to isolated goat corneal epithelial cells separately were assessed. The result shows the enzyme digestion method was more suitable for isolation and culture of limbal epithelial cells than explant culture. Isolated Limbal epithelial cells gained by enzyme digestion method appear polygonal shapes and better proliferation and these cells can proliferate for 20 passages in vitro. Significant difference of morphology and proliferating capacity between pre-cryopreserved corneal epithelial cells and the ones after resuscitation was not found. The results of Immunostaining and RT-PCR showed that the cultured cells expressed some phenotype-characteristics of corneal epithelial cells and limbal stem cell, for instance, K3/K12, p63, Cx43, ABCG2 and PCNA. Therefore these isolated cells were made up of limbal epithelial cells and limbal stem cells and could be employed to rebuild tissue engineering corneal epithelium.
2. Reconstruction of tissue engineering corneal epithelium
Limbal explants and cultured limbal epithelial cells of Guanzhong dairy goats were seeded on the human denuded amniotic membrane for reconstructing tissue engineering corneal epithelium, whose histological morphology, ultrastructure and immunocytochemiacal characteristic were observed. Results showed that limbal stem cell could proliferate on amniotic membrane and form 6～7 layers of stratified epithelium. The cells of stratified epithelium contained abundant organelles and glycongenosomes, and there were a lot of desmosomes and hemidemosome between cells or cells and amnioticmembrane. Immunostaining of paraffinic slice for p63 was positive. Comparing the growth performance of limbal epithelial cells from limbal explants seeded respectively on uncryo-preserved, cryo-preserved 30-day, cryo-preserved 90-day, cryo-preserved 180-day human amniotic membrane which had been removed of epithelium, we concluded that the cellular outgrowing area from limbal explants seeded on cryo-preserved 30-day human amniotic membrane was biggest, the cellular outgrowing area from limbal explants seeded on fresh human amniotic membrane was smallest. The growth performance of limbal epithelial cells seeded on cryo-preserved 60-day human amniotic membrane removed of epithelium and unremoved of epithelium was compared, the results showed that the cellular outgrowing area from limbal explants seeded on denuded amniotic membrane were significantly larger than that of intact amniotic membrane and implicated that both limbal explants and cultured limbal epithelial cells seeded on denuded human amniotic membrane can be used to reconstruct tissue engineering epithelium, the 30-day denuded amniotic membrane were most suitable to growth of limbal epithelial cells. The results provided evidences for studying tissue engineering artificial cornea.
3. Isolation, Culture and Identification of Goat Corneal Endothelial Cells
Four methods including explant culture, trypsin digestion, dispase digestion and dispase with trypsin digestion method were employed to isolate goat corneal endothelial cells (GCECs)in this experiment. The effectiveness of the methods to isolate GCECs was assessed. It was an optimum method to isolate GCECs in vitro using dispase with trypsin digestion. The obtained cells by the method could be cultured for about 20-passage. The resuscitated cryo-preserved GCECs performed the same morphology and proliferating capacity as that of cryo-preserved GCECs. Resorting to scan and transmission electron microscope, we saw that the morphology of isolated GCECs resembled that of GCECs in vivo. The cultured GCECs in vitro were measured by immunocytochemical staining and RT-PCR, of which results showed that the cells expressed NSE andβ3-tublin of neuronal marker, GFAP of astroglial cell marker, ZO-1 and Cx43 of cell connecting protein marker,but not expressed Nestin of neuronal marker, p63 of limbal stem cell marker, K3/K12 of corneal epithelial cell marker. The influence of different substances coating plastic ware,Ⅳcollagen, gelatin and poly-L-lysine, on the adherence and proliferation of GCECs was assessed in these experiments. It was found thatⅣcollagen , gelatin and poly-L-lysine greatly improved the adherence and proliferation of GCECs.Ⅳcollagen and gelatin could promote GCECs adherence and proliferation more remarkably than poly-L-lysine, but there was no distinct difference betweenⅣcollagen and gelatin. While gelatin is less expensive thanⅣcollagen and more appropriate for GCECs culture in vitro. Basing on the experiment about optimizing culture medium for culturing GCECs, we have obtained following conclusions. Both FBS and NBS can promote cell proliferation and maintain cellular morphology, but FBS was more appropriate. Although bFGF could also promote cell proliferation remarkably, it could also change the morphology of GCECs into fibroblast-like. EGF could not promote cell proliferation, while also influenced cellular morphology. Chondroitin sulfate could inhibit cell proliferation and maintain cellular morphology. Vitamin C could not influence cell proliferation and cellular morphology. By optical microscope, scan and transmission electron microscope, we could conclude that cultured GCECs in vitro were seeded on the human denuded amniotic membrane could form a monoptychial tissue on amniotic membrane. The result also implicated the isolated GCECs had the characteristics of native GCECs and could be cultured in vitro to form a monoptychial on amniotic membrane. The cells can be used for reconstrcting tissue engineering epithelium.
4. Transfection of hTERT gene in GCECs
The human telomerase reverse transcriptase(hTERT) gene was transferred into GCECs. The results of immunocytochemical staining and RT-PCR showed that hTERT gene was expressed in GCECs, which could proliferate for 20 passages after being transfected by hTERT gene. But there was no significant difference of cellular proliferation between transfected cells and untransfected cells. The study implicated that the proliferation of GCECs could not be promoted by transfecting hTERT gene into GCECs.
引文
[1] 李凤鸣.眼科全书[M].北京:人民卫生出版社,1996.
[2] 谢立信,董晓光.角膜移植学[M].北京:人民卫生出版社,2000.
[3] 肖仁度.实用眼科解剖学[M].太原:山西人民出版社,1980.
[4] 杨朝忠.现代角膜移植学[M].北京:人民军医出版社,1998.
[5] 刘家琦,李凤鸣.实用眼科学[M].北京:人民卫生出版社(第二版),2005.
[6] Dua H, Azuara-Blanco A. Limbal stem cells of the corneal epithelium[J].Survey of Ophthalmology,2000,44(5):415-425.
[7] 裴雪涛.干细胞生物学[M].北京:科学出版社,2003.
[8] Davanger M, Evensen A. Role of the pericorneal papillary structure in renewal of corneal epithelium[J].Nature,1971,229:560-561.
[9] Schemer A, Galvin S, Sun T T. Differentiation-related expression of a major 64K corneal keratin in vivo and in culture cells[J].J Cell Biol,1986,103:49-62.
[10] Cotsarelis G, Cheng S Z, Dong G, et al. Existence of slowcycling limbal epithelial basal cells that can be preferentially stimulated to proliferate:implications on epithelial stem cells[J].Cell,1989,57:201-209.
[11] Kurpakus M A, Stock E I, Jones J C. Expression of the 55-kD/64kD corneal keratins in ocular surface epithelium[J].Investigative Ophthalmology & Visual Science, 1990,31: 448-456.
[12] Ebato B, Friend J, Thoft R A. Comparision of limbal and peripheral human corneal epithelium in tissure culture[J].Investigative Ophthalmology & Visual Science,1988, 29:1533-1537.
[13] Lindberg K, Brown M E, Chaves H V, et al. In vitro propagation of human ocular surface epithelial cells for transplantation[J].Investigative Ophthalmology & Visual Science, 1993,34:2672-2679.
[14] Chen J J, Tseng S C. Abnormal corneal epithelial wound healing in partial-thickness removal of limbal epithelium[J].Investigative Ophthalmology & Visual Science,1991,32: 2219-2233.
[15] Huang A J, Tseng S C. Corneal epithelial wound healing in the absence of limbal epithelium[J].Investigative Ophthalmology & Visual Science,1991,32:96-105.
[16] Puangsricharern V, Tseng S C. Cytological evidence of corneal diseases with limbal stem cell deficiency[J].Ophthalmology,1995,102:1476-1485.
[17] Kenyon K R, Tseng S C. Limbal autograft transplantation for ocular surface disorder[J]. Ophthalmology,1989,96(6):709-723.
[18] Tsai R J, Sun T T, Tseng S C. Comparision of limbal and conjunctival autograft transplantation in corneal surface reconstruction in rabbits[J].Ophthalmology,1990, 97:446-455.
[19] Sharma A, Coles W H. Kinetics of corneal epithelial maintenance and graft loss. A population balance model[J].Investigative Ophthalmology & Visual Science,1989,30: 1962-1971.
[20] Hogan M J, Alvarado J A, Weddell J E. Histology of the human eye[M].Philadelphia: Saunders W B Co,1971,112-126.
[21] Cotsarelis G, Cheng S Z, Dong G, et al. Existence of slowcycling limbal epithelial basal cells that can be preferentially stimulated to proliferate: implications on epithelial stem cells[J].Cell,1989,57:201-209.
[22] Lavker R M, Dong G, Cheng S Z, et al. Relative proliferative rates of limbal and corneal epithelial. Implications of corneal epithelial migration, circadian rhythm, and suprabasally located DNA-synthesizing keratinocytes[J].Investigative Ophthalmology & Visual Science,1991,32:1864-1875.
[23] Gipson I K. The epithelial basement membrane zone of the limbus[J].Eye,1989,3:132-140.
[24] Zieske J D. Perpetuation of stem cell in the eye[J].Eye,1994,8:163-169.
[25] Barros E J, Santos O F, Matsumoto K, et al. Differential tubulogenic and branching morphogenetic activities of growth factors:implications for epithelial tissue development[J].Proc Natl Acad Sci,1995,92:4412-4416.
[26] Morrison S J, Shan N M, Anderson D J. Regulatory mechanisms in stem cell biology[J]. Cell,1997,88:287-298.
[27] Tseng S C. Regulation and clinical implications of corneal epithelial stem cells[J]. Mol Biol Rep,1996,23:47-58.
[28] Watt F, Hogan A. Out of Eden:Stem cells and their niches[J].Science,2000 287,1427-1431.
[29] Fuchs E, Segre J A. Stem cells:A new lease on life[J].Cell,2000,100,143-155.
[30] Schermer A, Galvin S, Sun T T. Differentiation-related expression of a major 64K corneal keratin in vivo and in culture cells[J].J Cell Biol,1986,103:49-62.
[31] Chen Z, de Paiva C S, Luo L. Characterization of putative stem cell phenotype in human limbal epithelia[J].Stem Cell,2004,22:355-366.
[32] Hayashi K, Kenyon K R. Increased cytochrome oxidase activity in alkaliburned corneas[J]. Current Eye Research,1988,7:131-138.
[33] SundarRaj N, Kinchington P R, Wessel H, et al. A Rho-associated protein kinase: differentially distributed in limbal and corneal epithelia[J].Investigative Ophthalmology & Visual Science,1998,39:1266-1271.
[34] Anderson S C, SundarRaj N. Regulation of a rho-associated kinase expression during the corneal epithelial cell cycle[J].Investigative Ophthalmology & Visual Science, 2001,42:933-940.
[35] Bravo R, Frank R, Blundell P A, et al. Cyclin/PCNA is the auxiliary protein of DNA polymerase-delta[J].Nature,1987,326:515-517.
[36] Gan L, Fagerholm P, Ekenbark S. Expression of proliferating cell nuclear antigen in corneas kept in long term culture[J].Acta Ophthalmologic Scandinavica,1998,76:308- 313.
[37] Tseng S C, Li D Q. Comparison of protein kinase C subtypes expression between normal and aniridic human ocular surfaces:implications for limbal stem cell dysfunction in aniridia[J].Cornea,1996,15:168-178.
[38] Zhou S, Schuetz J D, Bunting K D, et al. The ABC transporter Bcrpl/ABCG2 is expressedin a wide variety of stem cells and is a molecular determinant of the side-population phenotype[J].Nat Med,2001,7:1028-1034.
[39] de Paiva C S, Chen Z, Corrales R M, et al. ABCG2 transporter identifies a population of clonogenic human limbal epithelial cells[J].Stem Cells,2005,23:63-73.
[40] Pellegrini G, Dellambra E, Golisano O, et al. p63 identifies keratinocyte stem cells[J]. Proc Acad Sci USA,2001,98:3156-3161.
[41] Arpitha P, Prajna N V, Srinivasan M, et al. High expression of p63 combined with a large N/C ratio defines a subset of human limbal epithelial cells: implications on epithelial stem cells[J].Investigative Ophthalmology & Visual Science,2005,46:3631- 3636.
[42] Dua H S, Joseph A, Shanmuganathan V A, et al. Stem cell differentiation and the effects of deficiency[J].Eye,2003,17:877-885.
[43] Tseng S C, Kruse F E, Merritt J, et al. Comparison between serum-free and fibroblast- cocultured single-cell clonal culture systems: evidence showing that epithelial anti-apoptotic activity is present in 3T3 fibroblast conditioned media[J].Current Eye Research,1996,15:973-984.
[44] 屈雷.组织工程角膜上皮与内皮的构建与移植[D].陕西杨凌:西北农林科技大学,2004.
[45] Kruse F E, Tseng S C. Differing regulation of proliferation of limbus and corneal epithelium caused by serum factors[J].Ophthalmologe,1993,90(6):669-678.
[46] Kruse F E , Tseng S C. Serum differentially modulates the clonal growth and differentiation of cultured limbal and corneal epithelium[J].Investigative ophthalmology & visual,1993,34(10):2976-2989.
[47] 史伟云,董晓光,郭萍,等.兔角膜缘干细胞的体外培养[J].眼科研究,2002,2:114- 116.
[48] Wilson S E, He Y G, Weng J, et al. Effect of epidermal growth factor, hepatocyte growth factor, and keratinocyte growth factor, on proliferation, motility and differentiation of human corneal epithelial cells[J].Experimental Eye Research,1994, 59:665-678.
[49] Kruse F E,Tseng S C. Retinoic acid regulates clonal growth and differentiation of cultured limbal and peripheral corneal epithelium[J].Investigative Ophthalmology & Visual Science,1994,35:2405-2420.
[50] Kruse F E,Tseng S C. Proliferative and differentiative response of corneal and limbal epithelium to extracellular calcium in serum-free clonal cultures[J].J Cell Physiol, 1992,151(2):347-360.
[51] Wilson S E, He Y G, Lloyd S A. EGF,EGF receptor, basic FGF, TGF beta-1, and IL-1 alpha mRNA in human corneal epithelial cells and stromal fibroblasts[J].Investigative Ophthalmology & Visual Science,1992,33:1756-1765.
[52] Tamiya S, Delamere N A. Studies of Tyrosine Phosphorylation and Src Family Tyrosine Kinases in the Lens Epithelium[J].Investigative Ophthalmology & Visual Science,2005, 46(6):2076-2081.
[53] Zieske J D, Mason V S, Wasson M E, et al. Basement membrane assembly and differentiation of cultured corneal cells:importance of culture environment and endothelial cellinteraction[J].Experimental cell research,1994,214(2):621-633.
[54] 张杰,刘伟.角质干细胞及其在皮肤组织工程中的应用前景[J].中国实用美容整形外科,2004, 15(1):40-43.
[55] Pellegrini G, Golisano O, Paterna P, et al. Location and clonal analysis of stem cells and their differentiated progeny in the human ocular surface[J].J Cell Biol,1999, 145(4):769-782.
[56] Joseph A, Shanmuganathan V, McElveen J, et al. Limbal stem cells-Do we have a marker?[J]. Investigative Ophthalmology & Visual Science,2003,44:2029.
[57] Salehi-had H, Alvarenga L, Schwab I R, et al. Expression of p63 in Cultured Corneal Epithelium is a Function of Cell Confluence[ J].Investigative Ophthalmology & Visual Science,2004,45:3771.
[58] Yeh L K, Tsai R J. p63 stained the transit amplifying cells and not the marker for stem cells[J].Investigative Ophthalmology & Visual Science,2003,44:1353.
[59] Watanabe K, Nishida K. Yamato M, et al. Human limbal epithelium contains side population cells expressing the ATP-binding cassette transporter ABCG2[J].FEBS Letters,2004, 565(1-3):6-10.
[60] Matic M, Evans W H, Brink P R, et al. Epidermal stem cell do not communicate through gap junctions[J].Journal of Investigative Dermatology,2002,118(1):110-116.
[61] 陈晓东.表皮干细胞表面标记物的研究进展[J].国外医学(生理、病理科学与临床分册), 2004, 24(2):202-204.
[62] Chen Z, Evans W H, Pfluqfelder S C, et al. Gap junction protein connexin43 serves as a negative marker for a stem cell-containing population of human limbal epithelial cells[J].Stem Cells,2006,24(5):1265-1273.
[63] Kim H, Jun S X, de Paiva C S, et al. Phenotypic characterization of human corneal epithelial cells expanded ex vivo from limbal explant and single cell cultures[J]. Experimental Eye Research,2004,79(1):41-49.
[64] Chen J Y, Tseng S C. Corneal epithelial wound healing in partial limbal deficiency[J]. Investigative Ophthalmology & Visual Science,1990,31:1301-1314.
[65] Tsai R J, Tseng S C. Human allograft limbal transplantation for corneal surface reconstruction[J].Cornea,1994,13(5):389-400.
[66] Tsubota K, Satake Y, Kaido M, et al. Treatment of severe ocular-surface disorders with corneal epithelial stem cell transplantation[J].New England Journal of Medicine,1999, 340:1697-1703.
[67] Daya S M,Ilari F A. Living related conjunctival limbal allograft for the treatment of stem cell deficiency[J].Ophthalmology,2001,108(1):126-133.
[68] 郑利民,高莉莉,辛向阳.角膜缘干细胞移植的研究进展[J].内蒙古医学杂志,2006,27:641- 644.
[69] Pellegrini G,Traverso C E,Franzq A T, et al. Long-term restoration of damaged corneal surfaces with autologous cultivated corneal epithelium[J].Lancet,1997,349(7):990- 993.
[70] Tsai R J,Li L M,Cheng J K. Reconstruction of damaged corneas by transplantation ofautologous limbal epithelial cells[J].New England Journal of Medicine,2000,343(2): 86-93.
[71] Williams K A, Brereton H M, Aggarwal R, et al. Use of DNA polymorphisms and the polymerase chain reaction to examine the survival of a human limbal stem cell allograft[J].American journal of ophthalmology,1995,120(3):342-350.
[72] 刘春民,邓宏伟,周风,等.培养的同墓因免角膜缘干细胞移植治疗兔眼表烧伤[J].国际眼科杂志,2006,6:1309-1311.
[73] 潘志强,张文华.培养角膜缘干细胞羊膜移植片移植治疗角膜缘功能障碍[J].中国实用眼科杂志,2000,18(9):524-526.
[74] Koizumi N, Inatomi T,Suzuki T,et al. Cultivated corneal epithelial stem cell transplantation in ocular surface disorder[J].Ophthalmology,2001,108(9):1569-1574.
[75] Davis J W. Skin transplantation with a review of 550 cases at the Johns Hopkins Hospital[J].Johns Hopkins Medical Journal,1910,15:307-396.
[76] de Rotth A. Plastic repair of conjunctival defects with fetal membrane[J].Arch Ophthalmo1,1940,23:522-525.
[77] Kim J C,Tseng S C. Transplantation of preserved human amniotic membrane for surface reconstruction in severely damaged rabbits cornea[J].Cornea,1995,14(5):473-484.
[78] 郑世靖.羊膜在眼科领域应用的基础及临床研究进展[J].国外医学眼科学分册,2002,26(6): 344-348.
[79] Akle C A, Adinolfi M,Wesh K I,et al. Immunogenicity of human amniotic epithelial cells after transplantation into volunteers[J].Lancet,1981,2(8254):1003-1005.
[80] Grueterich M, Espana E, Tseng S C. Connexin 43 expression and proliferation of human limbal epithelium on intact and denuded amniotic membrane[J].Investigative Ophthalmology & Visual Science,2002,43(1):63-71.
[81] Abraham S,Mark R, Dagoberto M,et al. Suppression of interleukin laand interleukin 10 in human limbal epithelial cells cultured on the amniotic membrane stromal matrix[J]. British Journal of Ophthalmology,2001,85(4):444-449.
[82] Tseng S C, Li D Q, Ma X. Surppression of TGFβl,β2,β3,and TGF-βreceptor II expression and myofibroblast differentiation in human corneal and limbal fibroblasts by amniotic membrane matrix[J].J Cell Physio1,1999,179(3):325-335.
[83] Tsubota K, Toda I,Saito H ,et al. Reconstruction of the corneal epithelium by 1imbal allograft transplantation for severe ocular surface disorders[J].Ophthalmology,1995, 102:1486-1496.
[84] Tseng S C, Prabhasawat P ,Lee S H. Amniotic membrane transplantation for conjunctival surface reconstruction[J].American Journal of Ophthalmology,1997,124:765-774.
[85] 刘绍辉,王传富,赵桂秋,等.羊膜保存方法及其对羊膜活性影响的研究[J].中国实用眼科杂志, 2002,20(11):812-817.
[86] Nakamura T, Yosbitani M. Sterilized, freeze-dried Amniotic membrane: a useful substrate for ocular surface reconstruction[J].Investigative Ophthalmology & Visual Science,2004,45(1):93-99.
[87] 钟一声,叶纹,沈玺,等.冷冻保存羊膜的超微结构观察[J].眼科学报,2001,17(4):202-205.
[88] Madhavan H N, Priya K, Malathi J, et a1.Preparation of amniotic membrane for ocular surface reconstruction[J].Indian J Ophthalmol,2002,50(3):227-231.
[89] Shimazaki J,Shinozaki N, Tsubota K. Transplatation of amniotic membrane and limbral autograft for patients with recurrent pterygium associated with symblepharon[J]. British Journal of Ophthalmology,1998,82(3):235-240.
[90] Koizumi N, Inatomi T, Quantock A, et al. Amniotic membrane as a substrate for cultivating limbal corneal epithelial cells for autologous transplantation in rabbits[J].Cornea,2000,19(1):65-71.
[91] Gris O, Campo Z,Wolley-Dod C, et al. Amniotic membrane transplatation as a theraputic contact lens for the treatment of epithelial disorders [J].Cornea,2002,21(1):22-27.
[92] Solomon A, Meller D, Prabhasawat P,et al. Amniotic membrane grafts for nontraumatic corneal perforations, descemetoceles and deep ulcers[J].Ophthalmology,2002,109(4): 694-703.
[93] Thoft R A. Conjunctival transplantation[J].Arch Ophthalmology,1977,95(8): 1425-1427.
[94] Kwitko S, Marinho D, Barcaro S ,et a1. Allograft conjunctival transplantation for bilateral ocular surface disorders[J].Ophthalmology,1995,102(7):1020-1025.
[95] Shore J W ,Foster C S,Westfall C T, et al . Results of buccal mucosal grafting for patents with medically controlled ocular cicatricial pemphigoid[J].Ophthalmology, 1992,99(3):383-395.
[96] Levin P S, Button J J. Polytef ( Polytetrafluoroethylene) alloplastic grafting as a substitue for mucous membrane[J].Arch Ophthalmol,1990,108(2):282-285.
[97] Naumann G O, Lang, G K, Rummely V, et al .Autologous nasal mucosa transplantation in severe bilateral conjunctival mucous deficiency syndrome[J].ophthalmology,1990, 97(8):l0ll-1017.
[98] Barabino S,Rolando M, Bentivoglio G, et al. Role of amniotic membrane transplantation for conjunctival reconstruction in ocular cicatricial pemphigoid[J].Ophthalmology, 2003,110(3):474-480.
[99] Kobayashi A, Takahira M, Yamada A. Fornix and conjunctiva reconstruction by amniotic membrane in a patient with conjunctival mucosa-associated lymphoid tissue lymphoma[J].Japanese Journal of Ophthalmology,2002,46(3):346-348.
[100] 陈炜,刘祖国.羊膜移植治疗大泡性角膜病变[J].中国实用眼科杂志,2002,20(4):297.
[101] Mejia L F,Santamaria J P,Acosta C. Symptomatic management of postoperative bullous keratopathy with nonpreserved human amniotic membrane[J].Cornea,2002,21(4):342- 345.
[102] Pires R T,Chokshi A, Tseng S C. Amniotic membrane transplantation or conjunctival limbal autograft for limbal stem cell deficiency induced by 5-fluorouracil in glaucoma surgeries[J].Cornea,2000,19(3):284-287.
[103] Fujishima H,Shimazaki J,Shinazaki N, et al. Trabeculectomy with the use of amniotic membrane for uncontrollable glaucoma[J].Ophthalmic Surg Lasers,1998,29(5):428-431.
[104] 谢立信.第四届国际眼表和泪液大会概述[J].中华眼科杂志,2000,36(1):77-78.
[105] Kim H S, Sah W J,Kim Y J,et al. Amniotic membrane, tear film, corneal,and aqueouslevels of ofloxacin in rabbit eyes after amniotic membrane transplantation[J].Cornea, 2001,20(6):628.
[106] 闰国和,粟永萍,艾国平.人羊膜负载猪角朊细胞重建表皮的形态学研究[J].第三军医大学学报,2004,24(8):933-936.
[107] 张仁礼,李海标.胚胎干细胞源表皮样干细胞分化潜能的初步研究[J].解剖学报,2004, 35(1):65-68.
[108] 闰国和,粟永萍,艾国平,等.羊膜促进间充质干细胞表皮细胞生长的形态学特征[J].中国临床康复,2002,6(6):814-815.
[109] 陈燕图,黄仁胜,蔡永林,等.羊膜敷料在烧伤创面的临床应用[J].华夏医学,2004,17(1):44- 45.
[110] Murphy C, Alvarado J, Juster R, et al. Prenatal and postnatal cellularity of the human corneal endothelium:A quantitative histologic study[J].Investigative Ophthalmology & Visual Science,1984,25:312-322.
[111] Ko M K, Park W K, Lee J H, et al. A histomorphometric study of corneal endothelial cells in normal human fetuses[J].Experimental Eye Research,2001,72(4):403-409.
[112] Hollingsworth J, Perez-Gomez I, Mutalib H A, et al. A population study of the normal cornea using an in vivo slit scanning confocal microscope[J].Investigative Optometry & Vision Science,2001,78(10):706-711.
[113] Cho K S, Lee E H, Choi J S, et al. Reactive oxygen species-induced apoptosis and necrosis in bovine corneal endothelial cells[J].Investigative Ophthalmology & Visual Science,1999,40(5):911-919.
[114] Koh S W, Waschek J A. Corneal endothelial cell survival in organ cultures under acute oxidative stress: effect of VIP[J].Investigative Ophthalmology & Visual Science, 2000,41(13):4085-4092.
[115] Edelhauser H F. The resiliency of the corneal endothelium to refractive and intraocular surgery[J].Cornea,2000,19(3):263-273.
[116] Chung E H, Hutcheon A E, Joyce N C, et al. Synchronization of the G1-S transition in response to corneal debridement[J].Investigative Ophthalmology & Visual Science, 1999,40(9):1952-1958.
[117] Zieske J D, Takahashi H, Hutcheon A E, et al. Activation of epidermal growth factor receptor during corneal epithelial migration[J].Investigative Ophthalmology & Visual Science,2000,41(6):1346-1355.
[118] Laing R A, Neubauer L, Oak S S, et al. Evidence for mitosis in the adult corneal endothelium[J].Ophthalmology,1984,91:1129-1134.
[119] Matsubara M, Tanishima T. Wound-healing of corneal endothelium in monkey: an autoradiographic study[J].Japanese Journal of Ophthalmology,1983,27(3):444-450.
[120] Sherrard E S. The corneal endothelium in vivo: its response to mild trauma[J]. Experimental Eye Research,1976,22(4):347-357.
[121] Honda H, Ogita Y, Higuchi S, et al. Cell movements in a living mammalian tissue: long-term observation of individual cells in wounded corneal endothelia of cats[J]. Journal of Morphology,1982,174(1):25-39.
[122] Kaufman H E, Katz J I. Pathology of the corneal endothelium[J].Investigative Ophthalmology & Visual Science,1977,16:265-268.
[123] Joyce N C, Meklir B, Neufeld A H. In vitro pharmacologic separation of corneal endothelial migration and spreading responses[J].Investigative Ophthalmology & Visual Science,1990,31:1816-1826.
[124] Ichijima H, Petroll W M, Jester J V, et al. In vivo confocal microscopic studies of endothelial wound healing in rabbit cornea[J].Cornea,1993,12(5):369-378.
[125] Petroll W M, Jester J V, Barry-Lane P, et al. Assessment of f-actin organization and apical-basal polarity during in vivo cat endothelial wound healing[J].Investigative Ophthalmology & Visual Science,1995,36:2492-2502.
[126] Gordon S R, Buxar R M. Inhibition of cytoskeletal reorganization stimulates actin and tubulin syntheses during injuryinduced cell migration in the corneal endothelium[J].Journal of Cellular Biochemistry,1997,67(3):409-421.
[127] 李轩,崔立飞,田钟秀.人表皮生长因子对体外培养兔角膜内皮细胞影响的实验研究[J].中国实用眼科杂志,2000,18:161-163.
[128] Crawford K M, Ernst S A, Meyer R F, et al. NaK-ATPase pump sites in cultured bovine corneal endothelium of varying cell density at confluence[J].Investigative Ophthalmology & Visual Science,1995,36:1317-1326.
[129] Petroll W M, Jester J V, Bean J, et al. Labeling of cycling corneal endothelial cells during healing with a monoclonal antibody to the Ki67 antigen (MIB-1)[J].Cornea,1999, 18(1):98-108.
[130] Van G B, Fagerholm P, Philipson B, et al. Growth factors and their receptors in the anterior chamber. Absence of epidermal growth factor and transforming growth factor alpha in human aqueous humor[J].Ophthalmic Research,1996,28:361-364.
[131] Schulz M W, Chamberlain C G, De R U, et al . Acidic and basic FGF in ocular media and lens: implicationsfor lens polarity and growth patterns[J].Development,1993, 118(1):117-126.
[132] Araki-Sasaki K, Danjo S, Kawaguchi S, et al. Human hepatocyte growth factor (HGF) in the aqueous humor[J].Japanese Journal of Ophthalmology,1997,41:409-413.
[133] Welge-Lussen U, May C A, Neubauer A S, et al. Role of tissue growth factor in aqueous humor homeostasis[J].Current Opinion in Ophthalmology,2001,12(2):94-99.
[134] Wilson S E,Walker J W ,Chwang E L, et at. Hepatocyte growth factor,keratinocyte growth factor,their receptors,fibroblast growth factor receptor2 and the cell of the cornea[J].Investigative Ophthalmology & Visual Science,1993,34:2544.
[135] 潘红卫,赵长松.角膜内皮细胞增殖的研究进展[J].安用医学进修杂,2002,3O:5-8.
[136] Joyce N C,Harris D L,Mello D M.Mechanisms of mitotic inhibition in corneal endothelium:contact inhibition and TGF-beta2[J].Investigative Ophthalmology & Visual Science,2002,43(7):2152-2159.
[137] Senoo T, Obara Y, Joyce N C. EDTA:promotes proliferation in human corneal endothelium[J].Investigative Ophthalmology & Visual Science,2000,41:2930-2935.
[138] Joyce N C, Harris D L, Zieske J D, et al. Mitotic inhibition of corneal endotheliumin neonatal rats[J].Investigative Ophthalmology & Visual Science,1998,39:2572- 2583.
[139] Joyce N C, Meklir B, Joyce S J, et al. Cell cycle protein expression and proliferative status in human corneal cells[J].Investigative Ophthalmology & Visual Science, 1996, 37(4):645-655.
[140] Bednarz J.Doubilei V,Wollnik P C,et a1.Effect of three different media on serum free CUhure of donor corneas and isolated human corneal endothelial cells[J].British Journal of Ophthalmology,2001,85(12):1416-1420..
[141] Shin J S, Jang I K, Kim C W, et al. Development and characterization of a rabbit corneal endothelial cell line[J].Japanese Journal of Ophthalmology,2004,48(5):454-459.
[142] 樊廷俊,赵君,付永锋,等.家兔角膜内皮细胞系的建立[J].中国科学 C 辑:生命科学,2007, 37:20-27.
[143] Ikebe H, Takamatsu T, Itoi M, et al. Cytofluorometric nuclear DNA determinations on human corneal endothelial cells[J].Experimental Eye Research,1984,39:497-504.
[144] Ikebe H, Takamatsu T, Itoi M, et al. Age-dependent changes in nuclear DNA content and cell size of presumably normal human corneal endothelium[J].Experimental Eye Research,1986,43,251-258.
[145] Ikebe H, Takamatsu T, Itoi M, et al. Changes in nuclearDNA content and cell size of injured human corneal endothelium[J].Experimental Eye Research,1988,47:205-215.
[146] Wilson S E, Lloyd S A. Epidermal growth factor and its receptor, basic fibroblast growth factor, transforming growth factor-beta1 and interleukin-1 alpha messenger RNA production in human corneal endothelial cells[J].Investigative Ophthalmology & Visual Science,1991,32:2747-2756.
[147] Samples J R, Binder P S, Nayak S K, et al. Propagation of human corneal endothelium: in vitro effect of growth factors[J].Experimental Eye Research,1991,52:121-128.
[148] Hyldahl L. Primary cell cultures from human embryonic corneas[J].Journal of Cell Science,1984,66:343-351.
[149] Insler M S, Lopez J G. Microcarrier cell culture of neonatal human corneal endothelium[J].Current Eye Research,1990,9:23-30.
[150] Nayak S K, Binder P S. The growth of endothelium from human corneal rims in tissue culture[J].Investigative Ophthalmology & Visual Science,1984,25,1213-1216.
[151] Simonsen A H, Sorensen K E, Sperling S, et al. Thymidine incorporation by human corneal endothelium during organ culture[J].Acta Ophthalmol(Copenh),1981,59:110-118.
[152] Senoo T,Joyce N C.Cell cycle kinetics in corneal endothelium from old and young donors[J].Investigative Ophthalmology & Visual Science,2000,41(3):660-667..
[153] Chan K Y, Lindquist T D, Edenfield M J,et al.Pharmacokinetic study of recombinant human epidermal growth factor in the anterior eye[J].Investigative Ophthalmology & Visual Science,1991,32(13):3209-3215.
[154] Woost P G, Tumblatt M M, Eifeman R A, et al. Growth factors and corneal endothelial cells DNA synthesis by defined growth fectoxs[J].Cornea,1992,14:1-5.
[155] Johnstone E W,Wong H C,Coster D J,et a1.Factors affecting bovine corneal endothelialcell density in vitro[J].British Journal of Ophthalmology,1996,8O(3):256-262.
[156] 郁熙明.血小板生长因子的活性和性质[J].国外医学(内科学分册),1988,15(2):49-53.
[157] Kamiyama K, Iguchi I, Wang X,et al. Enhancement of growth of rabbit corneal endothelial cells by PDGF[J].Cornea,1995,14:187-195.
[158] Woost P G, Jumblatt M M, Eiferman R A, et al. Growth factors and corneal endothelial cells: II. Characterization of epidermal growth factor receptor from bovinecorneal endothelial cells[J].Cornea,1992,11:11-19.
[159] Iguchi I, Kamiyama K, Wang X, et al. Enhancing effect of platelet-derived growth factors on migration of corneal endothelial cells[J].Cornea,1995,14:365-371.
[160] Lev M R. Nerve growth factor years later[J].Science,l987,237:1154.
[161] Lambiase A, Manni L, Bonini S, et al. Nerve growth factor promotes corneal healing:structural, biochemical and molecular analyses of rat and human cornea[J]. Investigative Ophthalmology & Visual Science,2000,41:1063-1069.
[162] 李中国,邱良秀,赵长松,等.神经生长因子对角膜内皮细胞增殖的影响[J].眼科研究,2001, 19:312-314.
[163] 钟一声.生长因子与角膜内皮细胞[J].眼科研究,1999,17:314-316.
[164] Chen K H, Azar D,Joyce N C.Transplantation of adult human corneal endothelium ex vivo:a morphologic study[J].Cornea,2001,20(7):731-737..
[165] 李姝燕,吴静,郝念,等.层粘连蛋白和表皮生长因子对兔角膜内皮细胞周期的影响[J].眼科新进展,2004,24:9-11.
[166] Blake D A,Yu H,Young D L,et a1.Matrix stimulates the proliferation of human corneal endothelial cells in culture[J].Investigative Ophthalmology & Visual Science,1997, 38(6):1119-1129..
[167] Schimmelpfennig B H. Direct and indirect determination of nonuniform cell density distribution in human corneal endothelium[J].Investigative Ophthalmology & Visual Science,1984,25:223-229.
[168] Amann J, Holley G P, Lee S B, et al. Increased endothelial cell density in the paracentral and peripheral regions of the human cornea[J].American Journal of Ophthalmology,2003,135:584-590.
[169] Bednarz J, Rodokanaki-von Schrenck A, Engelmann K. Different characteristics of endothelial cells from central and peripheral human cornea in primary culture and after subculture[J].In Vitro Cell Dev Biol Anim,1998,34:149-153.
[170] Paull A C, Whikehart D R. Expression of the p53 family of proteins in central and peripheral human corneal endothelial cells[J].Mol Vis,2005,11:328-334.
[171] Mimura T, Yamagami S, Yokoo S, et al. Comparison of Rabbit Corneal Endothelial Cell Precursors in the Central and Peripheral Cornea[J].Investigative Ophthalmology & Visual Science,2005,46:3645-3648.
[172] Yokoo S, Yamagami S, Yanagi Y, et al. Human Corneal Endothelial Cell Precursors Isolated by Sphere-Forming Assay[J].Investigative Ophthalmology & Visual Science, 2005,46:1626-1631.
[173] Mimura T, Yamagami S, Yokoo S, et al. Sphere Therapy for Corneal Endothelium Deficiencyin a Rabbit Model[J].Investigative Ophthalmology & Visual Science,2005,46:3128- 3135.
[174] Whikehart D R, Parikh C H, Vaughn A V, et al. Evidence suggesting the existence of stem cells for the human corneal endothelium[J].Molecular Vision,2005,11:816-824.
[175] Engelmann K, B?hnke M, Friedl P, et al. Isolation and long-term cultivation of human corneal endothelial cells[J].Investigative Ophthalmology & Visual Science,1988, 29:1656-1662.
[176] Neukld A H,Jumblatt M M, Matkin E D, et al. Maintenance of corneal endothelial cell shape by prostaglandin E2: effects of EGF and indomethacin[J].Investigative Ophthalmology & Visual Science,1986,27:1437-1442.
[177] Engelmann K,Bednarz J,Schafer HJ,et a1.Isolation and characterization of a mouse monoclonal antibody against human corneal endothelial cells[J]Experimental Eye Research,2001,73(1):9-16.
[178] Hayashi K, Sueishi K, Tanaka K, et al. Immunohistochemical evidence of the origin of human endothelialcells and keratocytes[J]. Graefe's Archive for Clinical and Experimental Ophthalmology,1986,224:452-456.
[179] Michetti F, Rende M, Lauriola L, et al. Immunohistochemical localization of S100-containing cells in non-nervous structures of the human eye[J].Cell Biology International Reports,1986,10:765-773.
[180] Yagihashi S, Kimura M, Kurotaki H, et al. Gastric submucosal tumors of neurogenic origin with neuroaxonal and Schwann cell elements[J].American Journal of Pathology,1987,153:41-50.
[181] B?hnke M, Vogelberg K, Engelmann K. Detection of neurone-specific enolase in long-term cultures of human corneal endothelium[J].Graefe's Archive for Clinical and Experimental Ophthalmology,1998,236:522-526.
[182] Jumblatt M M,Maurice D M,McCulley J P. Transplantation of tissue cultured corneal endothelium[J].Investigative Ophthalmology & Visual Science,1978,17:1135-1141.
[183] Lange T M,Wood T O,MeLaughlin B J. Corneal endothelial cell transplantation using Descemet’s membrane as carrier[J].Journal of Cataract and Refractive Surgery,1993, 19:232-235.
[184] Sbiro Amano.Transplantation of corneal endothelial cells[J].Nippon Ganka Gakkai Zasshi,2002,106(12):805-835.
[185] Ishino Y,Sano Y,Nakamura T,et al. Amniotic emembrane as a Carrier for cultivated human corneal endothelial cells transplantation[J]. Investigative Ophthalmology & Visual Science,2004,45:800-806.
[186] 曹谊林,汪铮,李晓红,等.组织工程研究中的胚胎干细胞问题[J].世界科技研究与发展,2000, 22(6):54-58.
[187] 翁雨来,商庆新,曹谊林.生命科学的新增长点--组织工程[J].牙体牙髓牙周病学杂志,2000, 10(5):249-254.
[188] Elsdale T, Bard J. Collagen substrate for studies on cell behavior[J].Journal of Cell Biology,1972,54(3):626-637.
[189] Minami Y, Sugihara H, Oono S. Reconstruction of cornea in three-dimensional collagen gel matrix culture[J].Investigative Ophthalmology & Visual Science,1993,34(7):2316- 2324.
[190] Griffith M, Osborne R, Xiong X, et al. Functional human corneal equivalents constructed from cell lines[J].Science,1999,286(5447):2169-2172.
[191] 陈家祺,张胜,郭琳洁.体外培养重建角膜组织的技术探讨[J].中国实用眼科杂志,2001, 19(11):815-817.
[192] Lindberg K, Brown M E, Chaves H V, et al. In vitro preparation of human ocular surface epithelial cells for transplantation[J].Investigative Ophthalmology & Visual Science,1993,34:2672-2679.
[193] Kinoshita S. Ocular surface reconstruction by tissue engineering[J].Nippon Ganka Gakkai Zasshi,2002,106(12):837-868.
[194] Ray J T, Li L M, Chen J K. Reconstruction of Damaged Corneas by Transplantation of Autologous Limbal Epithelial Cells[J].New England Journal of Medicine,2000,343:86- 93.
[195] Behrouz A, Harry M. Germ cells frommouse and human embryonic stem cells[J]. Reproduction,2006,132(5):699-707.
[196] Rambhatla L, Chiu C P, Kundu P, et al. Generation of hepatocyte-like cells fromhuman embryonic stem cells[J].Cell Transplant,2003,12(1):1-11.
[197] Maxim A, Vodyanik L, Jack A, et al. Human embryonic stem cell-derived CD34 cells: efficient production in the co-culture with OP9 stromal cells and analysis of lymphohematopoietic potential[J].Blood,2005,105(2):617- 626.
[198] Reubinoff B E, Itsykson P. Neural progenitors from human stem cells[J].Nature Biotechnology,2001,19(12):1134-1140.
[199] Kwon Y D, Oh S K, Kim H S, et al. Cellular manipulation of human embryonic stem cells by TAT-PDX1 protein transduction[J].Molecular Therapy,2005,12(1):28-32.
[200] Ahmad S, Stewart R, Yung S,et al. Differentiation of Human Embryonic Stem Cells into Corneal Epithelial-Like Cells by In Vitro Replication of the Corneal Epithelial Stem Cell Niche[J].Stem Cells,2007,25(5):1145-1155.
[201] 黄冰,王智崇,葛坚,等.利用皮肤干细胞横向分化重建角膜上皮的初步研究[J].中华医学杂志, 2004,84:838-842.
[202] Yu L, Ge J, Wang Z C, et al. Preliminary experimental study of induced Diferentiation of embryonic stem cells into corneal epithelialcells[J].Eye Science,2001,17:138- 143.
[203] Inatomi T, Nakamura T, Koizumi N, et al. Midterm results on ocular surface reconstruction using cultivated autologous oral mucosal epithelial transplantation [J].American Journal of Ophthalmology,2006,141(2):267-275.
[204] Araki-Sasaki K, Ohashi Y, Sasabe T, et al. An SV40-immortalized human corneal epithelial cell line and its characterization[J].Investigative Ophthalmology & Visual Science,1995,36:614-621.
[205] Jin Y, Liu Y, Zhang C, et al. Treatment of rabbit cornea wounds with skin epidermalstem cells[J].Acta Academiae Medicinae Sinicae,2005,27(6):674-677.
[206] 彭亮红,柳林.角膜缘干细胞标志物[J].国际眼科杂志,2005,5(5):1032-1036.
[207] Hayashida Y, Nishida K, Yamato M, et al. Ocular surface reconstruction using autologous rabbit oralmucosalepithelial sheets fabricated ex vivo on a temperature-responsive culture surface[J].Investigative Ophthalmology & Visual Science,2005,46(5):1632-1639.
[208] Hata K, Kagami H, Ueda M, et al. The characteristics of cultured mucosal cell sheetasa material for grafting:comparisonwith cultured epidermalcell sheet[J].Annals of Plastic Surgery,1995,34(5):530-538.
[209] Ueda M, Hata K, Horie K, et al. The potentialofor almucosal cells for cultured epithelium: a prelim inary report[J].Annals of Plastic Surgery,1995,35(5):498-504.
[210] Nakamura T, Endo K, Cooper L J, et al. The successful culture and autologous transplantation of rabbit oralmucosal epithelial cells on amniotic membrane[J]. Investigative Ophthalmology & Visual Science,2003,44(1):106-116.
[211] Nishida K, Yamato M, Hayashida Y, et al. Corneal reconstruction with tissue-engineered cell sheets composed ofautologous oral mucosal epithelium[J].New England Journal of Medicine,2004,351(12):1187-1196.
[212] Insler M S, Lopez J G. Heterologous transplantation verus enhance-ment of human corneal endothelium[J].Cornea,1991,10:136-148.
[213] Peters D M, Dowd N, Brandt C, et al. Human papilloma virus E6/E7 genes can expand the lifespan of human corneal fibroblasts[J].In Vitro Cellular & Developmental Biology-Animal,1996,32(5):279-284.
[214] Reichl S, Bednarz J, Muller-Goymann C C. Human corneal equivalent as cell culture model for in vitro drug permeation studies[J].British Journal of Ophthalmology, 2004,88(4):560-565.
[215] Aboalchamat B, Engelmann K, Bohnke M, et al. Morphological and functional analysis of immortalized human corneal endothelial cells after transplantation[J]. Experimental Eye Research,1999,69(5):547-553.
[216] Joyce N C. Proliferative capacity of the corneal endothelium[J].Progress in Retinal and Eye Research,2003,22:359-389.
[217] 易伟斌,胡竹林,徐岩泽,等.羊膜作为载体培养血管内皮细胞替代自体角膜内皮细胞的研究[J].昆明医学院学报,2006,27:1-4.
[218] Engelmann K, Bednarz J, Valtink M. Prospects for endothelial transplantation[J]. Experimental Eye Research,2004,78:573-578.
[219] 丁勇,徐锦堂,陈建苏.组织工程角膜材料研究进展[J].中国病理生理杂志,2002,18:1557- 1560.
[220] Koizumi N, Inatomi T, Sotozono C, et al. Growth factor mRNAandprotein in preserved human amnioticmembrane[J].Current Eye Research,2000,20:173- 177.
[221] Hao Y, Ma D K, Hwang D G, et al.Identification of antiangiogenic and anti inflammatory proteins in human amniotic membrane[J].Cornea,2000,19:348- 352.
[222] Wang D Y, Hsueh Y J, Yang V C, et al. Propagation and Phenotypic Preservation of RabbitLimbal Epithelial Cells on Amniotic Membrane[J].Investigative Ophthalmology & Visual Science,2003,44:4698-4705.
[223] Espana E M, He H, Kawakita T, et al. Human keratocytes cultured on amniotic membrane stroma preserve morphology and express keratocan [J].Investigative Ophthalmology & Visual Science,2003,44(12):5136- 5141.
[224] He Y G, Alizadeh H, Kinoshita K, et al. Experimental transplantation of cultured human limbal and amniotic epitheial cells onto the corneal surface[J].Cornea,1999,18:570- 579.
[225] Mimura T, Yamagami S, Yokoo S, et al. Cultured human corneal endothelial cell transplantation with a collagen sheet in a rabbit model[J].Investigative Ophthalmology & Visual Science,2004,45:2992-2997.
[226] Ebato B, Friend J, Thoft R A. Comparison of central and peripheral human corneal epithelium in tissue culture[J].Investigative Ophthalmology & Visual Science,1987, 28:1450.
[227] 吴静,徐锦堂,赵松滨.培养角膜上皮移植的实验研究[J].中国实用眼科杂志.2001,19:178- 179.
[228] Engelmann K, Drexler D, B?hnke M. Transplantation of adult human or porcine corneal endothelial cells onto human recipients in vitro.Part I:Cell culturing and transplantation procedure[J].Cornea,1999,18:199-206.
[229] B?hnke M, Eggli P, Engelmann K. Transplantation of cultured adult human or porcine endothelial cells onto human recipients in vitro.Part II: Evaluation in the scanning electron microscope[J].Cornea,1999,18:207-213.
[230] Jumblatt M M, Maurice D M, Schwartz B D. A gelatin membrane substrate for the transplantation of tissue cultured cells[J].Transplantation,1980,29: 498-499.
[231] Mohay J, Lange T M, Soltau J B, et al. Transplantation of corneal endothelial cell using a cell carrier device[J].Cornea,1994,13:173-182.
[232] 郭萍,谢立信,董晓光.人角膜缘干细胞在体外不同载体上培养的实验研究[J].眼科研究, 2001,19(4):304-307.
[233] Hadlock T, Singh S, Vacanti J P, et al. Ocular cell monolayers cultured on biodegradable substrates[J].Tissue Eng,1999,5:187-196.
[234] Griffith M, Hakim M, Shimmura S, et al. Artificial human corneas Scaffolds for transplantation and host regeneration[J].Cornea,2002,21(2):54-61.
[235] Shinazaki J, Aiba M, Goto E, et al. Transplantation of human limbal epithelium cultivated on amniotic membrane for the treatment of severe ocular surface disorders[J].Ophthalmology,2002,109:1285-1290.
[236] 顾国贞,白靓,王芳,等.兔角膜缘干细胞体外分离、培养和生物学鉴定的实验研究[J].现代生物医学进展,2007,7:533-536.
[237] 郭萍,史伟云,李绍伟,等.人角膜缘干细胞体外培养后生物学特性的变化[J].眼科新进展, 2002,22:229-231.
[238] 张晓敏,孙慧敏,赵少贞,等.兔角膜缘干细胞两种培养方法的比较[J].眼科研究,2005,23: 575-578.
[239] Stepp M A, Zhu L. Upregulation of alpha 9 integrin and tenascin during epithelial regeneration after debridement in the cornea[J].J Histochem cytochem,1997,45: 189-201.
[240] Hernandez G E E, Theiss C, Steuhl K P, et al. Expression of p63 in response to phorbol ester in human limbal epithelial cells expanded on intact human amniotic membrane[J]. Investigative Ophthalmology & Visual Science,2003,44:2959-2965.
[241] Matic M, Petrov I N, Chen S, et al. Stem cells of the corneal epithelium lack connexins and metabolite transfer capacity[J].Differentiation,1997,61:251-260.
[242] Harkin DG, Barnard Z, Gillies P, et al. Analysis of p63 and cytokeratin expression in a cultivated limbal autograft used in the treatment of limbal stem cell deficiency [J].Br J Ophthalmol,2004,88:1154-1158.
[243] Djalilian A R, Mahesh S P, Koch C A, et al. Survival of Donor Epithelial Cells after Limbal Stem Cell Transplantation[J].Investigative Ophthalmology and Visual Science, 2005,46:803-807.
[244] Du Y, Chen J, Funderburgh J L, et al. Functional reconstruction of rabbit corneal epithelium by human limbal cells cultur ed on amniotic membrane[J].Mol Vis,2003,9: 635-643.
[245] Kim H, Oh Y. The Effect of Limbal Transplantation Versus Amniotic Membrane Transplantation on Limbal Damaged Rabbit Cornea [J].Investigative Ophthalmology & Visual Science,2003,44:904.
[246] Sangwan V S, Matalia H P, Vemuganti G K, et al. Early Results of Penetrating Keratoplasty After Cultivated Limbal Epithelium Transplantation[J].Arch Ophthalmol, 2005,123(3):334-340.
[247] Yeh L K, Chen W L, Li W, et al. Soluble Lumican Glycoprotein Purified from Human Amniotic Membrane Promotes Corneal Epithelial Wound Healing[J].Investigative Ophthalmology & Visual Science,2005,46(2):479-486.
[248] Li H C, Niederkorn J Y, Neelam S, et al. Immunosuppressive Factors Secreted by Human Amniotic Epithelial Cells[J].Investigative Ophthalmology & Visual Science,2005, 46(3):900-907.
[249] Avila M, Espana M, Moreno C, et al. Reconstruction of ocular surface with heterologous limbal epithelium and amniotic membrane in a rabbit model[J].Cornea,2001,20(4):414- 420.
[250] Grueterich M, Espana E M, Tseng S C. Ex vivo expansion of limbal epithelial stem cells: Amniotic membrane serving as a stem cell niche[J].Surv Ophthalmol,2003,48(6):631- 646.
[251] Nakamura T, Koizumi N, Tsuzuki M, et al. Successful regrafting of cultivated corneal epithelium using amniotic membrane as a carrier in severe ocular surface disease[J]. Cornea,2003,22(1):70-71.
[252] Ramaesh K, Dhillon B. Ex vivo expansion of corneal limbal epithelial/stem cells for corneal surface reconstruction[J].Eur J Ophthalmol,2003,13(6):515-524.
[253] Kubo M,Sonada Y,Muramatsu R,Usui M.Immunogenicity of human amniotic membrane inexperimental xenotransplantation[J].Investigative Ophthalmology & Visual Science, 2001,42:1539-1546.
[254] Barfort P, Maurice D. Electrical potential and fluid transport across the corneal endothelium[J].Experimental Eye Research,1974,19:11-19.
[255] Maurice D M. The location of the fluid pump in the cornea[J].The Journal of Physiology, 1972,221:43-54.
[256] Jun A S, Chakravarti S, Edelhauser H F, et al. Aging changes of mouse corneal endothelium and descemet’s membrane[J].Experimental Eye Research,2006,83:890-896.
[257] Slick W C, Mannagh J and Yuhasz Z. Enzymatic removal and pure culture of rabbit corneal endothelial cells[J].Archives of Ophthalmology,1965,73:229-232.
[258] Engelmann K, Friedl P. Growth of human corneal endothelial cells in a serum- reduced medium[J].Cornea,1995,14:62-70.
[259] Diane A B, Haini Y, Danna L Y,et al. Matrix stimulates the proliferation of human corneal endothelial cells in culture[J].Investigative Ophthalmology & Visual Science,1997,38:1119-1129.
[260] Kazunori M, Jonathan D, Yasuhiro O, et al. Effect of donor age on morphologic variation of cultured human corneal endothelial cells[J].Cornea,2001,20:59-63.
[261] Pan T, Zhao J, Fu Y, et al. Establishment of a novel corneal endothelial cell line from domestic rabbit, Oryctolagus curiculus[J].Science in China Series C:Life Sciences,2007,50:161-169.
[262] Joo C K, Green W R, Pepose J S, et al. Repopulation of denuded murine descemet’s membrane with life-extended murine corneal endothelial cells as a model for corneal cell transplantation[J].Graefe's Archives for Clinical and Experimental Ophthalmology,2000,238:174-180.
[263] Savion N, Isaacs J D, Shuman M A, et al. Proliferation and differentiation of bovine corneal endothelial cells in culture[J].Metabolic Pediatric Systemic Ophthalmology, 1982,6:305-320.
[264] Ogita Y, Nakamura T, Higuchi S, et al. Histochemical studies of mitochondrial activities of cultured corneal endothelial cells of cat during wound-healing[J]. Japanese Journal of Ophthalmology,1990,34:200-215.
[265] Koizumi N, Sakamoto Y, Okumura N, et al. Cultivated corneal endothelial cell sheet transplantation in a primate model [J].Investigative Ophthalmology & Visual Science, 2007,48:4519-4526.
[266] Cintia S P, Zhuo C, Rosa M C, et al. ABCG2 transporter identifies a population of clonogenic human limbal epithelial cells[J].Stem Cell,2005,23:63-73.
[267] Konomi K, Joyce N C. Age and topographical comparison of telomere lengths in human corneal endothelial cells[J].Molecular Vision,2007,13:1251-1258.
[268] Suh L H, Zhang C, Chuck R, et al. Cryopreservation and lentiviral-mediated genetic modification of human primary cultured corneal endothelial cells[J].Investigative Ophthalmology & Visual Science,2007,48:3056-3061.
[269] Stocker F W, Eiring A, Georgiade R, et al. A tissue culturetechnique for growingcorneal epithelial, stromal,and endothelialtissues separately[J].Am J Ophthalmol, 1958,46:294-298.
[270] Zhu C C, Joyce N C. Proliferative response of corneal endothelial cells from young and older donors[J].Investigative Ophthalmology & Visual Science,2004,45:1743- 1751.
[271] Joyce N C, Zhu C C. Human corneal endothelial cell proliferation: potential for use in regenerative medicine[J].Cornea,2004,23:S8-S19.
[272] Woost P G, Jumblatt M M, Eiferman R A, et al. Growth factors and corneal endothelial cells: I. Stimulation of bovine corneal endothelial cell DNA synthesis by defined growth factors[J].Cornea,1992:11:1-10.
[273] Raymond G M, Jumblatt M M, Bartels S P, et al. Rabbit corneal endothelial cells in vitro: effects of EGF[J].Investigative Ophthalmology & Visual Science,1986,27: 474-479.
[274] Nayak S K, Samples J R, Deg J K, et al. Growth characteristics of primate (baboon) corneal endothelium in vitro[J].Investigative Ophthalmology & Visual Science,1986,27:607-611.
[275] Germain L, Auger F A. Tissue engineered biomaterials: biological and mechanical characteristics. Encyclopedic Handbook of Biomaterials and Bioengineering,part B; Applications[M].New York,USA:Marcel Dekker,inc.Publishers,1995,699-734.
[276] Yue BY, Niedra R, Baum J L. Effects of ascorbic acid on cultured rabbit corneal endothelial cells[J].Investigative Ophthalmology & Visual Science,1980,19:1471- 1476.
[277] Wilson S E, Weng J, Blai S, et al. Expression of E6/E7 or SV40 Large T Antigen- Coding Oncogenes in Human Corneal Endothelial Cells Indicates Regulated High-Proliferative Capacity[J].Investigativc Ophthalmology & Visual Science,1995,36:32-40.
[278] Minamino T, Komuro I.Role of telomere in endothelial dysfunction in atherosclerosis[J].Curr Opin Lipidol,2002,13:537-543.
[279] 谢晓繁,贾赤宇,陈璧,等.端粒酶活性对人表皮干细胞分化影响的实验研究[J].实用医学杂志,2006,22:1480-1482.
[280] Murasawa S, Llevadot J, Silver M, et al. Constitutive human telomerase reverse transcriptase expression enhances regenera2 Live properties of endothelial progenitor cells[J].Circulation,2002,106:1133-1139.
[281] Takamasa W, Norikuni S, Westerman K A, et al. Establishment of immortalized human hepatic stellate scavenger cells to develop bioartificial livers[J].Transplantation 2003,75:1873-1880.
[282] Wege H, Le H T, Chui M S, et al. Telomerase reconstition immortalizes human fetal hepatocytes without disrupting their differentiation potential[J].Gastroenterology, 2003,124:432-444.
[283] Turnpenny L, Brickwood S, Spalluto C M, et al. Derivation of Human Embryonic Germ Cells:An alternative source of pluripotent stem cells[J].Stem Cells,2003,21:598- 609.
[284] Bekaert S, De Meyer T, Van O P. Telomere attrition as ageing biomarker[J]. Anticancer Res,2005,25:3011-3021.
[285] Egan C A, Savre-Train I, Shay J W, et al. Analysis of Telomere Lengths in Human Corneal Endothelial Cells from Donors of Different Ages[J].Investigative Ophthalmology & Visual Science,1998,39:648-653.
[286] Whikehart D R, Register S J, Chang Q, et al. Relationship of Telomeres and p53 in Aging Bovine Corneal Endothelial Cell Cultures[J].Investigative Ophthalmology & Visual Science,2000,41:1070-1075.
[2] 谢立信,董晓光.角膜移植学[M].北京:人民卫生出版社,2000.
[3] 肖仁度.实用眼科解剖学[M].太原:山西人民出版社,1980.
[4] 杨朝忠.现代角膜移植学[M].北京:人民军医出版社,1998.
[5] 刘家琦,李凤鸣.实用眼科学[M].北京:人民卫生出版社(第二版),2005.
[6] Dua H, Azuara-Blanco A. Limbal stem cells of the corneal epithelium[J].Survey of Ophthalmology,2000,44(5):415-425.
[7] 裴雪涛.干细胞生物学[M].北京:科学出版社,2003.
[8] Davanger M, Evensen A. Role of the pericorneal papillary structure in renewal of corneal epithelium[J].Nature,1971,229:560-561.
[9] Schemer A, Galvin S, Sun T T. Differentiation-related expression of a major 64K corneal keratin in vivo and in culture cells[J].J Cell Biol,1986,103:49-62.
[10] Cotsarelis G, Cheng S Z, Dong G, et al. Existence of slowcycling limbal epithelial basal cells that can be preferentially stimulated to proliferate:implications on epithelial stem cells[J].Cell,1989,57:201-209.
[11] Kurpakus M A, Stock E I, Jones J C. Expression of the 55-kD/64kD corneal keratins in ocular surface epithelium[J].Investigative Ophthalmology & Visual Science, 1990,31: 448-456.
[12] Ebato B, Friend J, Thoft R A. Comparision of limbal and peripheral human corneal epithelium in tissure culture[J].Investigative Ophthalmology & Visual Science,1988, 29:1533-1537.
[13] Lindberg K, Brown M E, Chaves H V, et al. In vitro propagation of human ocular surface epithelial cells for transplantation[J].Investigative Ophthalmology & Visual Science, 1993,34:2672-2679.
[14] Chen J J, Tseng S C. Abnormal corneal epithelial wound healing in partial-thickness removal of limbal epithelium[J].Investigative Ophthalmology & Visual Science,1991,32: 2219-2233.
[15] Huang A J, Tseng S C. Corneal epithelial wound healing in the absence of limbal epithelium[J].Investigative Ophthalmology & Visual Science,1991,32:96-105.
[16] Puangsricharern V, Tseng S C. Cytological evidence of corneal diseases with limbal stem cell deficiency[J].Ophthalmology,1995,102:1476-1485.
[17] Kenyon K R, Tseng S C. Limbal autograft transplantation for ocular surface disorder[J]. Ophthalmology,1989,96(6):709-723.
[18] Tsai R J, Sun T T, Tseng S C. Comparision of limbal and conjunctival autograft transplantation in corneal surface reconstruction in rabbits[J].Ophthalmology,1990, 97:446-455.
[19] Sharma A, Coles W H. Kinetics of corneal epithelial maintenance and graft loss. A population balance model[J].Investigative Ophthalmology & Visual Science,1989,30: 1962-1971.
[20] Hogan M J, Alvarado J A, Weddell J E. Histology of the human eye[M].Philadelphia: Saunders W B Co,1971,112-126.
[21] Cotsarelis G, Cheng S Z, Dong G, et al. Existence of slowcycling limbal epithelial basal cells that can be preferentially stimulated to proliferate: implications on epithelial stem cells[J].Cell,1989,57:201-209.
[22] Lavker R M, Dong G, Cheng S Z, et al. Relative proliferative rates of limbal and corneal epithelial. Implications of corneal epithelial migration, circadian rhythm, and suprabasally located DNA-synthesizing keratinocytes[J].Investigative Ophthalmology & Visual Science,1991,32:1864-1875.
[23] Gipson I K. The epithelial basement membrane zone of the limbus[J].Eye,1989,3:132-140.
[24] Zieske J D. Perpetuation of stem cell in the eye[J].Eye,1994,8:163-169.
[25] Barros E J, Santos O F, Matsumoto K, et al. Differential tubulogenic and branching morphogenetic activities of growth factors:implications for epithelial tissue development[J].Proc Natl Acad Sci,1995,92:4412-4416.
[26] Morrison S J, Shan N M, Anderson D J. Regulatory mechanisms in stem cell biology[J]. Cell,1997,88:287-298.
[27] Tseng S C. Regulation and clinical implications of corneal epithelial stem cells[J]. Mol Biol Rep,1996,23:47-58.
[28] Watt F, Hogan A. Out of Eden:Stem cells and their niches[J].Science,2000 287,1427-1431.
[29] Fuchs E, Segre J A. Stem cells:A new lease on life[J].Cell,2000,100,143-155.
[30] Schermer A, Galvin S, Sun T T. Differentiation-related expression of a major 64K corneal keratin in vivo and in culture cells[J].J Cell Biol,1986,103:49-62.
[31] Chen Z, de Paiva C S, Luo L. Characterization of putative stem cell phenotype in human limbal epithelia[J].Stem Cell,2004,22:355-366.
[32] Hayashi K, Kenyon K R. Increased cytochrome oxidase activity in alkaliburned corneas[J]. Current Eye Research,1988,7:131-138.
[33] SundarRaj N, Kinchington P R, Wessel H, et al. A Rho-associated protein kinase: differentially distributed in limbal and corneal epithelia[J].Investigative Ophthalmology & Visual Science,1998,39:1266-1271.
[34] Anderson S C, SundarRaj N. Regulation of a rho-associated kinase expression during the corneal epithelial cell cycle[J].Investigative Ophthalmology & Visual Science, 2001,42:933-940.
[35] Bravo R, Frank R, Blundell P A, et al. Cyclin/PCNA is the auxiliary protein of DNA polymerase-delta[J].Nature,1987,326:515-517.
[36] Gan L, Fagerholm P, Ekenbark S. Expression of proliferating cell nuclear antigen in corneas kept in long term culture[J].Acta Ophthalmologic Scandinavica,1998,76:308- 313.
[37] Tseng S C, Li D Q. Comparison of protein kinase C subtypes expression between normal and aniridic human ocular surfaces:implications for limbal stem cell dysfunction in aniridia[J].Cornea,1996,15:168-178.
[38] Zhou S, Schuetz J D, Bunting K D, et al. The ABC transporter Bcrpl/ABCG2 is expressedin a wide variety of stem cells and is a molecular determinant of the side-population phenotype[J].Nat Med,2001,7:1028-1034.
[39] de Paiva C S, Chen Z, Corrales R M, et al. ABCG2 transporter identifies a population of clonogenic human limbal epithelial cells[J].Stem Cells,2005,23:63-73.
[40] Pellegrini G, Dellambra E, Golisano O, et al. p63 identifies keratinocyte stem cells[J]. Proc Acad Sci USA,2001,98:3156-3161.
[41] Arpitha P, Prajna N V, Srinivasan M, et al. High expression of p63 combined with a large N/C ratio defines a subset of human limbal epithelial cells: implications on epithelial stem cells[J].Investigative Ophthalmology & Visual Science,2005,46:3631- 3636.
[42] Dua H S, Joseph A, Shanmuganathan V A, et al. Stem cell differentiation and the effects of deficiency[J].Eye,2003,17:877-885.
[43] Tseng S C, Kruse F E, Merritt J, et al. Comparison between serum-free and fibroblast- cocultured single-cell clonal culture systems: evidence showing that epithelial anti-apoptotic activity is present in 3T3 fibroblast conditioned media[J].Current Eye Research,1996,15:973-984.
[44] 屈雷.组织工程角膜上皮与内皮的构建与移植[D].陕西杨凌:西北农林科技大学,2004.
[45] Kruse F E, Tseng S C. Differing regulation of proliferation of limbus and corneal epithelium caused by serum factors[J].Ophthalmologe,1993,90(6):669-678.
[46] Kruse F E , Tseng S C. Serum differentially modulates the clonal growth and differentiation of cultured limbal and corneal epithelium[J].Investigative ophthalmology & visual,1993,34(10):2976-2989.
[47] 史伟云,董晓光,郭萍,等.兔角膜缘干细胞的体外培养[J].眼科研究,2002,2:114- 116.
[48] Wilson S E, He Y G, Weng J, et al. Effect of epidermal growth factor, hepatocyte growth factor, and keratinocyte growth factor, on proliferation, motility and differentiation of human corneal epithelial cells[J].Experimental Eye Research,1994, 59:665-678.
[49] Kruse F E,Tseng S C. Retinoic acid regulates clonal growth and differentiation of cultured limbal and peripheral corneal epithelium[J].Investigative Ophthalmology & Visual Science,1994,35:2405-2420.
[50] Kruse F E,Tseng S C. Proliferative and differentiative response of corneal and limbal epithelium to extracellular calcium in serum-free clonal cultures[J].J Cell Physiol, 1992,151(2):347-360.
[51] Wilson S E, He Y G, Lloyd S A. EGF,EGF receptor, basic FGF, TGF beta-1, and IL-1 alpha mRNA in human corneal epithelial cells and stromal fibroblasts[J].Investigative Ophthalmology & Visual Science,1992,33:1756-1765.
[52] Tamiya S, Delamere N A. Studies of Tyrosine Phosphorylation and Src Family Tyrosine Kinases in the Lens Epithelium[J].Investigative Ophthalmology & Visual Science,2005, 46(6):2076-2081.
[53] Zieske J D, Mason V S, Wasson M E, et al. Basement membrane assembly and differentiation of cultured corneal cells:importance of culture environment and endothelial cellinteraction[J].Experimental cell research,1994,214(2):621-633.
[54] 张杰,刘伟.角质干细胞及其在皮肤组织工程中的应用前景[J].中国实用美容整形外科,2004, 15(1):40-43.
[55] Pellegrini G, Golisano O, Paterna P, et al. Location and clonal analysis of stem cells and their differentiated progeny in the human ocular surface[J].J Cell Biol,1999, 145(4):769-782.
[56] Joseph A, Shanmuganathan V, McElveen J, et al. Limbal stem cells-Do we have a marker?[J]. Investigative Ophthalmology & Visual Science,2003,44:2029.
[57] Salehi-had H, Alvarenga L, Schwab I R, et al. Expression of p63 in Cultured Corneal Epithelium is a Function of Cell Confluence[ J].Investigative Ophthalmology & Visual Science,2004,45:3771.
[58] Yeh L K, Tsai R J. p63 stained the transit amplifying cells and not the marker for stem cells[J].Investigative Ophthalmology & Visual Science,2003,44:1353.
[59] Watanabe K, Nishida K. Yamato M, et al. Human limbal epithelium contains side population cells expressing the ATP-binding cassette transporter ABCG2[J].FEBS Letters,2004, 565(1-3):6-10.
[60] Matic M, Evans W H, Brink P R, et al. Epidermal stem cell do not communicate through gap junctions[J].Journal of Investigative Dermatology,2002,118(1):110-116.
[61] 陈晓东.表皮干细胞表面标记物的研究进展[J].国外医学(生理、病理科学与临床分册), 2004, 24(2):202-204.
[62] Chen Z, Evans W H, Pfluqfelder S C, et al. Gap junction protein connexin43 serves as a negative marker for a stem cell-containing population of human limbal epithelial cells[J].Stem Cells,2006,24(5):1265-1273.
[63] Kim H, Jun S X, de Paiva C S, et al. Phenotypic characterization of human corneal epithelial cells expanded ex vivo from limbal explant and single cell cultures[J]. Experimental Eye Research,2004,79(1):41-49.
[64] Chen J Y, Tseng S C. Corneal epithelial wound healing in partial limbal deficiency[J]. Investigative Ophthalmology & Visual Science,1990,31:1301-1314.
[65] Tsai R J, Tseng S C. Human allograft limbal transplantation for corneal surface reconstruction[J].Cornea,1994,13(5):389-400.
[66] Tsubota K, Satake Y, Kaido M, et al. Treatment of severe ocular-surface disorders with corneal epithelial stem cell transplantation[J].New England Journal of Medicine,1999, 340:1697-1703.
[67] Daya S M,Ilari F A. Living related conjunctival limbal allograft for the treatment of stem cell deficiency[J].Ophthalmology,2001,108(1):126-133.
[68] 郑利民,高莉莉,辛向阳.角膜缘干细胞移植的研究进展[J].内蒙古医学杂志,2006,27:641- 644.
[69] Pellegrini G,Traverso C E,Franzq A T, et al. Long-term restoration of damaged corneal surfaces with autologous cultivated corneal epithelium[J].Lancet,1997,349(7):990- 993.
[70] Tsai R J,Li L M,Cheng J K. Reconstruction of damaged corneas by transplantation ofautologous limbal epithelial cells[J].New England Journal of Medicine,2000,343(2): 86-93.
[71] Williams K A, Brereton H M, Aggarwal R, et al. Use of DNA polymorphisms and the polymerase chain reaction to examine the survival of a human limbal stem cell allograft[J].American journal of ophthalmology,1995,120(3):342-350.
[72] 刘春民,邓宏伟,周风,等.培养的同墓因免角膜缘干细胞移植治疗兔眼表烧伤[J].国际眼科杂志,2006,6:1309-1311.
[73] 潘志强,张文华.培养角膜缘干细胞羊膜移植片移植治疗角膜缘功能障碍[J].中国实用眼科杂志,2000,18(9):524-526.
[74] Koizumi N, Inatomi T,Suzuki T,et al. Cultivated corneal epithelial stem cell transplantation in ocular surface disorder[J].Ophthalmology,2001,108(9):1569-1574.
[75] Davis J W. Skin transplantation with a review of 550 cases at the Johns Hopkins Hospital[J].Johns Hopkins Medical Journal,1910,15:307-396.
[76] de Rotth A. Plastic repair of conjunctival defects with fetal membrane[J].Arch Ophthalmo1,1940,23:522-525.
[77] Kim J C,Tseng S C. Transplantation of preserved human amniotic membrane for surface reconstruction in severely damaged rabbits cornea[J].Cornea,1995,14(5):473-484.
[78] 郑世靖.羊膜在眼科领域应用的基础及临床研究进展[J].国外医学眼科学分册,2002,26(6): 344-348.
[79] Akle C A, Adinolfi M,Wesh K I,et al. Immunogenicity of human amniotic epithelial cells after transplantation into volunteers[J].Lancet,1981,2(8254):1003-1005.
[80] Grueterich M, Espana E, Tseng S C. Connexin 43 expression and proliferation of human limbal epithelium on intact and denuded amniotic membrane[J].Investigative Ophthalmology & Visual Science,2002,43(1):63-71.
[81] Abraham S,Mark R, Dagoberto M,et al. Suppression of interleukin laand interleukin 10 in human limbal epithelial cells cultured on the amniotic membrane stromal matrix[J]. British Journal of Ophthalmology,2001,85(4):444-449.
[82] Tseng S C, Li D Q, Ma X. Surppression of TGFβl,β2,β3,and TGF-βreceptor II expression and myofibroblast differentiation in human corneal and limbal fibroblasts by amniotic membrane matrix[J].J Cell Physio1,1999,179(3):325-335.
[83] Tsubota K, Toda I,Saito H ,et al. Reconstruction of the corneal epithelium by 1imbal allograft transplantation for severe ocular surface disorders[J].Ophthalmology,1995, 102:1486-1496.
[84] Tseng S C, Prabhasawat P ,Lee S H. Amniotic membrane transplantation for conjunctival surface reconstruction[J].American Journal of Ophthalmology,1997,124:765-774.
[85] 刘绍辉,王传富,赵桂秋,等.羊膜保存方法及其对羊膜活性影响的研究[J].中国实用眼科杂志, 2002,20(11):812-817.
[86] Nakamura T, Yosbitani M. Sterilized, freeze-dried Amniotic membrane: a useful substrate for ocular surface reconstruction[J].Investigative Ophthalmology & Visual Science,2004,45(1):93-99.
[87] 钟一声,叶纹,沈玺,等.冷冻保存羊膜的超微结构观察[J].眼科学报,2001,17(4):202-205.
[88] Madhavan H N, Priya K, Malathi J, et a1.Preparation of amniotic membrane for ocular surface reconstruction[J].Indian J Ophthalmol,2002,50(3):227-231.
[89] Shimazaki J,Shinozaki N, Tsubota K. Transplatation of amniotic membrane and limbral autograft for patients with recurrent pterygium associated with symblepharon[J]. British Journal of Ophthalmology,1998,82(3):235-240.
[90] Koizumi N, Inatomi T, Quantock A, et al. Amniotic membrane as a substrate for cultivating limbal corneal epithelial cells for autologous transplantation in rabbits[J].Cornea,2000,19(1):65-71.
[91] Gris O, Campo Z,Wolley-Dod C, et al. Amniotic membrane transplatation as a theraputic contact lens for the treatment of epithelial disorders [J].Cornea,2002,21(1):22-27.
[92] Solomon A, Meller D, Prabhasawat P,et al. Amniotic membrane grafts for nontraumatic corneal perforations, descemetoceles and deep ulcers[J].Ophthalmology,2002,109(4): 694-703.
[93] Thoft R A. Conjunctival transplantation[J].Arch Ophthalmology,1977,95(8): 1425-1427.
[94] Kwitko S, Marinho D, Barcaro S ,et a1. Allograft conjunctival transplantation for bilateral ocular surface disorders[J].Ophthalmology,1995,102(7):1020-1025.
[95] Shore J W ,Foster C S,Westfall C T, et al . Results of buccal mucosal grafting for patents with medically controlled ocular cicatricial pemphigoid[J].Ophthalmology, 1992,99(3):383-395.
[96] Levin P S, Button J J. Polytef ( Polytetrafluoroethylene) alloplastic grafting as a substitue for mucous membrane[J].Arch Ophthalmol,1990,108(2):282-285.
[97] Naumann G O, Lang, G K, Rummely V, et al .Autologous nasal mucosa transplantation in severe bilateral conjunctival mucous deficiency syndrome[J].ophthalmology,1990, 97(8):l0ll-1017.
[98] Barabino S,Rolando M, Bentivoglio G, et al. Role of amniotic membrane transplantation for conjunctival reconstruction in ocular cicatricial pemphigoid[J].Ophthalmology, 2003,110(3):474-480.
[99] Kobayashi A, Takahira M, Yamada A. Fornix and conjunctiva reconstruction by amniotic membrane in a patient with conjunctival mucosa-associated lymphoid tissue lymphoma[J].Japanese Journal of Ophthalmology,2002,46(3):346-348.
[100] 陈炜,刘祖国.羊膜移植治疗大泡性角膜病变[J].中国实用眼科杂志,2002,20(4):297.
[101] Mejia L F,Santamaria J P,Acosta C. Symptomatic management of postoperative bullous keratopathy with nonpreserved human amniotic membrane[J].Cornea,2002,21(4):342- 345.
[102] Pires R T,Chokshi A, Tseng S C. Amniotic membrane transplantation or conjunctival limbal autograft for limbal stem cell deficiency induced by 5-fluorouracil in glaucoma surgeries[J].Cornea,2000,19(3):284-287.
[103] Fujishima H,Shimazaki J,Shinazaki N, et al. Trabeculectomy with the use of amniotic membrane for uncontrollable glaucoma[J].Ophthalmic Surg Lasers,1998,29(5):428-431.
[104] 谢立信.第四届国际眼表和泪液大会概述[J].中华眼科杂志,2000,36(1):77-78.
[105] Kim H S, Sah W J,Kim Y J,et al. Amniotic membrane, tear film, corneal,and aqueouslevels of ofloxacin in rabbit eyes after amniotic membrane transplantation[J].Cornea, 2001,20(6):628.
[106] 闰国和,粟永萍,艾国平.人羊膜负载猪角朊细胞重建表皮的形态学研究[J].第三军医大学学报,2004,24(8):933-936.
[107] 张仁礼,李海标.胚胎干细胞源表皮样干细胞分化潜能的初步研究[J].解剖学报,2004, 35(1):65-68.
[108] 闰国和,粟永萍,艾国平,等.羊膜促进间充质干细胞表皮细胞生长的形态学特征[J].中国临床康复,2002,6(6):814-815.
[109] 陈燕图,黄仁胜,蔡永林,等.羊膜敷料在烧伤创面的临床应用[J].华夏医学,2004,17(1):44- 45.
[110] Murphy C, Alvarado J, Juster R, et al. Prenatal and postnatal cellularity of the human corneal endothelium:A quantitative histologic study[J].Investigative Ophthalmology & Visual Science,1984,25:312-322.
[111] Ko M K, Park W K, Lee J H, et al. A histomorphometric study of corneal endothelial cells in normal human fetuses[J].Experimental Eye Research,2001,72(4):403-409.
[112] Hollingsworth J, Perez-Gomez I, Mutalib H A, et al. A population study of the normal cornea using an in vivo slit scanning confocal microscope[J].Investigative Optometry & Vision Science,2001,78(10):706-711.
[113] Cho K S, Lee E H, Choi J S, et al. Reactive oxygen species-induced apoptosis and necrosis in bovine corneal endothelial cells[J].Investigative Ophthalmology & Visual Science,1999,40(5):911-919.
[114] Koh S W, Waschek J A. Corneal endothelial cell survival in organ cultures under acute oxidative stress: effect of VIP[J].Investigative Ophthalmology & Visual Science, 2000,41(13):4085-4092.
[115] Edelhauser H F. The resiliency of the corneal endothelium to refractive and intraocular surgery[J].Cornea,2000,19(3):263-273.
[116] Chung E H, Hutcheon A E, Joyce N C, et al. Synchronization of the G1-S transition in response to corneal debridement[J].Investigative Ophthalmology & Visual Science, 1999,40(9):1952-1958.
[117] Zieske J D, Takahashi H, Hutcheon A E, et al. Activation of epidermal growth factor receptor during corneal epithelial migration[J].Investigative Ophthalmology & Visual Science,2000,41(6):1346-1355.
[118] Laing R A, Neubauer L, Oak S S, et al. Evidence for mitosis in the adult corneal endothelium[J].Ophthalmology,1984,91:1129-1134.
[119] Matsubara M, Tanishima T. Wound-healing of corneal endothelium in monkey: an autoradiographic study[J].Japanese Journal of Ophthalmology,1983,27(3):444-450.
[120] Sherrard E S. The corneal endothelium in vivo: its response to mild trauma[J]. Experimental Eye Research,1976,22(4):347-357.
[121] Honda H, Ogita Y, Higuchi S, et al. Cell movements in a living mammalian tissue: long-term observation of individual cells in wounded corneal endothelia of cats[J]. Journal of Morphology,1982,174(1):25-39.
[122] Kaufman H E, Katz J I. Pathology of the corneal endothelium[J].Investigative Ophthalmology & Visual Science,1977,16:265-268.
[123] Joyce N C, Meklir B, Neufeld A H. In vitro pharmacologic separation of corneal endothelial migration and spreading responses[J].Investigative Ophthalmology & Visual Science,1990,31:1816-1826.
[124] Ichijima H, Petroll W M, Jester J V, et al. In vivo confocal microscopic studies of endothelial wound healing in rabbit cornea[J].Cornea,1993,12(5):369-378.
[125] Petroll W M, Jester J V, Barry-Lane P, et al. Assessment of f-actin organization and apical-basal polarity during in vivo cat endothelial wound healing[J].Investigative Ophthalmology & Visual Science,1995,36:2492-2502.
[126] Gordon S R, Buxar R M. Inhibition of cytoskeletal reorganization stimulates actin and tubulin syntheses during injuryinduced cell migration in the corneal endothelium[J].Journal of Cellular Biochemistry,1997,67(3):409-421.
[127] 李轩,崔立飞,田钟秀.人表皮生长因子对体外培养兔角膜内皮细胞影响的实验研究[J].中国实用眼科杂志,2000,18:161-163.
[128] Crawford K M, Ernst S A, Meyer R F, et al. NaK-ATPase pump sites in cultured bovine corneal endothelium of varying cell density at confluence[J].Investigative Ophthalmology & Visual Science,1995,36:1317-1326.
[129] Petroll W M, Jester J V, Bean J, et al. Labeling of cycling corneal endothelial cells during healing with a monoclonal antibody to the Ki67 antigen (MIB-1)[J].Cornea,1999, 18(1):98-108.
[130] Van G B, Fagerholm P, Philipson B, et al. Growth factors and their receptors in the anterior chamber. Absence of epidermal growth factor and transforming growth factor alpha in human aqueous humor[J].Ophthalmic Research,1996,28:361-364.
[131] Schulz M W, Chamberlain C G, De R U, et al . Acidic and basic FGF in ocular media and lens: implicationsfor lens polarity and growth patterns[J].Development,1993, 118(1):117-126.
[132] Araki-Sasaki K, Danjo S, Kawaguchi S, et al. Human hepatocyte growth factor (HGF) in the aqueous humor[J].Japanese Journal of Ophthalmology,1997,41:409-413.
[133] Welge-Lussen U, May C A, Neubauer A S, et al. Role of tissue growth factor in aqueous humor homeostasis[J].Current Opinion in Ophthalmology,2001,12(2):94-99.
[134] Wilson S E,Walker J W ,Chwang E L, et at. Hepatocyte growth factor,keratinocyte growth factor,their receptors,fibroblast growth factor receptor2 and the cell of the cornea[J].Investigative Ophthalmology & Visual Science,1993,34:2544.
[135] 潘红卫,赵长松.角膜内皮细胞增殖的研究进展[J].安用医学进修杂,2002,3O:5-8.
[136] Joyce N C,Harris D L,Mello D M.Mechanisms of mitotic inhibition in corneal endothelium:contact inhibition and TGF-beta2[J].Investigative Ophthalmology & Visual Science,2002,43(7):2152-2159.
[137] Senoo T, Obara Y, Joyce N C. EDTA:promotes proliferation in human corneal endothelium[J].Investigative Ophthalmology & Visual Science,2000,41:2930-2935.
[138] Joyce N C, Harris D L, Zieske J D, et al. Mitotic inhibition of corneal endotheliumin neonatal rats[J].Investigative Ophthalmology & Visual Science,1998,39:2572- 2583.
[139] Joyce N C, Meklir B, Joyce S J, et al. Cell cycle protein expression and proliferative status in human corneal cells[J].Investigative Ophthalmology & Visual Science, 1996, 37(4):645-655.
[140] Bednarz J.Doubilei V,Wollnik P C,et a1.Effect of three different media on serum free CUhure of donor corneas and isolated human corneal endothelial cells[J].British Journal of Ophthalmology,2001,85(12):1416-1420..
[141] Shin J S, Jang I K, Kim C W, et al. Development and characterization of a rabbit corneal endothelial cell line[J].Japanese Journal of Ophthalmology,2004,48(5):454-459.
[142] 樊廷俊,赵君,付永锋,等.家兔角膜内皮细胞系的建立[J].中国科学 C 辑:生命科学,2007, 37:20-27.
[143] Ikebe H, Takamatsu T, Itoi M, et al. Cytofluorometric nuclear DNA determinations on human corneal endothelial cells[J].Experimental Eye Research,1984,39:497-504.
[144] Ikebe H, Takamatsu T, Itoi M, et al. Age-dependent changes in nuclear DNA content and cell size of presumably normal human corneal endothelium[J].Experimental Eye Research,1986,43,251-258.
[145] Ikebe H, Takamatsu T, Itoi M, et al. Changes in nuclearDNA content and cell size of injured human corneal endothelium[J].Experimental Eye Research,1988,47:205-215.
[146] Wilson S E, Lloyd S A. Epidermal growth factor and its receptor, basic fibroblast growth factor, transforming growth factor-beta1 and interleukin-1 alpha messenger RNA production in human corneal endothelial cells[J].Investigative Ophthalmology & Visual Science,1991,32:2747-2756.
[147] Samples J R, Binder P S, Nayak S K, et al. Propagation of human corneal endothelium: in vitro effect of growth factors[J].Experimental Eye Research,1991,52:121-128.
[148] Hyldahl L. Primary cell cultures from human embryonic corneas[J].Journal of Cell Science,1984,66:343-351.
[149] Insler M S, Lopez J G. Microcarrier cell culture of neonatal human corneal endothelium[J].Current Eye Research,1990,9:23-30.
[150] Nayak S K, Binder P S. The growth of endothelium from human corneal rims in tissue culture[J].Investigative Ophthalmology & Visual Science,1984,25,1213-1216.
[151] Simonsen A H, Sorensen K E, Sperling S, et al. Thymidine incorporation by human corneal endothelium during organ culture[J].Acta Ophthalmol(Copenh),1981,59:110-118.
[152] Senoo T,Joyce N C.Cell cycle kinetics in corneal endothelium from old and young donors[J].Investigative Ophthalmology & Visual Science,2000,41(3):660-667..
[153] Chan K Y, Lindquist T D, Edenfield M J,et al.Pharmacokinetic study of recombinant human epidermal growth factor in the anterior eye[J].Investigative Ophthalmology & Visual Science,1991,32(13):3209-3215.
[154] Woost P G, Tumblatt M M, Eifeman R A, et al. Growth factors and corneal endothelial cells DNA synthesis by defined growth fectoxs[J].Cornea,1992,14:1-5.
[155] Johnstone E W,Wong H C,Coster D J,et a1.Factors affecting bovine corneal endothelialcell density in vitro[J].British Journal of Ophthalmology,1996,8O(3):256-262.
[156] 郁熙明.血小板生长因子的活性和性质[J].国外医学(内科学分册),1988,15(2):49-53.
[157] Kamiyama K, Iguchi I, Wang X,et al. Enhancement of growth of rabbit corneal endothelial cells by PDGF[J].Cornea,1995,14:187-195.
[158] Woost P G, Jumblatt M M, Eiferman R A, et al. Growth factors and corneal endothelial cells: II. Characterization of epidermal growth factor receptor from bovinecorneal endothelial cells[J].Cornea,1992,11:11-19.
[159] Iguchi I, Kamiyama K, Wang X, et al. Enhancing effect of platelet-derived growth factors on migration of corneal endothelial cells[J].Cornea,1995,14:365-371.
[160] Lev M R. Nerve growth factor years later[J].Science,l987,237:1154.
[161] Lambiase A, Manni L, Bonini S, et al. Nerve growth factor promotes corneal healing:structural, biochemical and molecular analyses of rat and human cornea[J]. Investigative Ophthalmology & Visual Science,2000,41:1063-1069.
[162] 李中国,邱良秀,赵长松,等.神经生长因子对角膜内皮细胞增殖的影响[J].眼科研究,2001, 19:312-314.
[163] 钟一声.生长因子与角膜内皮细胞[J].眼科研究,1999,17:314-316.
[164] Chen K H, Azar D,Joyce N C.Transplantation of adult human corneal endothelium ex vivo:a morphologic study[J].Cornea,2001,20(7):731-737..
[165] 李姝燕,吴静,郝念,等.层粘连蛋白和表皮生长因子对兔角膜内皮细胞周期的影响[J].眼科新进展,2004,24:9-11.
[166] Blake D A,Yu H,Young D L,et a1.Matrix stimulates the proliferation of human corneal endothelial cells in culture[J].Investigative Ophthalmology & Visual Science,1997, 38(6):1119-1129..
[167] Schimmelpfennig B H. Direct and indirect determination of nonuniform cell density distribution in human corneal endothelium[J].Investigative Ophthalmology & Visual Science,1984,25:223-229.
[168] Amann J, Holley G P, Lee S B, et al. Increased endothelial cell density in the paracentral and peripheral regions of the human cornea[J].American Journal of Ophthalmology,2003,135:584-590.
[169] Bednarz J, Rodokanaki-von Schrenck A, Engelmann K. Different characteristics of endothelial cells from central and peripheral human cornea in primary culture and after subculture[J].In Vitro Cell Dev Biol Anim,1998,34:149-153.
[170] Paull A C, Whikehart D R. Expression of the p53 family of proteins in central and peripheral human corneal endothelial cells[J].Mol Vis,2005,11:328-334.
[171] Mimura T, Yamagami S, Yokoo S, et al. Comparison of Rabbit Corneal Endothelial Cell Precursors in the Central and Peripheral Cornea[J].Investigative Ophthalmology & Visual Science,2005,46:3645-3648.
[172] Yokoo S, Yamagami S, Yanagi Y, et al. Human Corneal Endothelial Cell Precursors Isolated by Sphere-Forming Assay[J].Investigative Ophthalmology & Visual Science, 2005,46:1626-1631.
[173] Mimura T, Yamagami S, Yokoo S, et al. Sphere Therapy for Corneal Endothelium Deficiencyin a Rabbit Model[J].Investigative Ophthalmology & Visual Science,2005,46:3128- 3135.
[174] Whikehart D R, Parikh C H, Vaughn A V, et al. Evidence suggesting the existence of stem cells for the human corneal endothelium[J].Molecular Vision,2005,11:816-824.
[175] Engelmann K, B?hnke M, Friedl P, et al. Isolation and long-term cultivation of human corneal endothelial cells[J].Investigative Ophthalmology & Visual Science,1988, 29:1656-1662.
[176] Neukld A H,Jumblatt M M, Matkin E D, et al. Maintenance of corneal endothelial cell shape by prostaglandin E2: effects of EGF and indomethacin[J].Investigative Ophthalmology & Visual Science,1986,27:1437-1442.
[177] Engelmann K,Bednarz J,Schafer HJ,et a1.Isolation and characterization of a mouse monoclonal antibody against human corneal endothelial cells[J]Experimental Eye Research,2001,73(1):9-16.
[178] Hayashi K, Sueishi K, Tanaka K, et al. Immunohistochemical evidence of the origin of human endothelialcells and keratocytes[J]. Graefe's Archive for Clinical and Experimental Ophthalmology,1986,224:452-456.
[179] Michetti F, Rende M, Lauriola L, et al. Immunohistochemical localization of S100-containing cells in non-nervous structures of the human eye[J].Cell Biology International Reports,1986,10:765-773.
[180] Yagihashi S, Kimura M, Kurotaki H, et al. Gastric submucosal tumors of neurogenic origin with neuroaxonal and Schwann cell elements[J].American Journal of Pathology,1987,153:41-50.
[181] B?hnke M, Vogelberg K, Engelmann K. Detection of neurone-specific enolase in long-term cultures of human corneal endothelium[J].Graefe's Archive for Clinical and Experimental Ophthalmology,1998,236:522-526.
[182] Jumblatt M M,Maurice D M,McCulley J P. Transplantation of tissue cultured corneal endothelium[J].Investigative Ophthalmology & Visual Science,1978,17:1135-1141.
[183] Lange T M,Wood T O,MeLaughlin B J. Corneal endothelial cell transplantation using Descemet’s membrane as carrier[J].Journal of Cataract and Refractive Surgery,1993, 19:232-235.
[184] Sbiro Amano.Transplantation of corneal endothelial cells[J].Nippon Ganka Gakkai Zasshi,2002,106(12):805-835.
[185] Ishino Y,Sano Y,Nakamura T,et al. Amniotic emembrane as a Carrier for cultivated human corneal endothelial cells transplantation[J]. Investigative Ophthalmology & Visual Science,2004,45:800-806.
[186] 曹谊林,汪铮,李晓红,等.组织工程研究中的胚胎干细胞问题[J].世界科技研究与发展,2000, 22(6):54-58.
[187] 翁雨来,商庆新,曹谊林.生命科学的新增长点--组织工程[J].牙体牙髓牙周病学杂志,2000, 10(5):249-254.
[188] Elsdale T, Bard J. Collagen substrate for studies on cell behavior[J].Journal of Cell Biology,1972,54(3):626-637.
[189] Minami Y, Sugihara H, Oono S. Reconstruction of cornea in three-dimensional collagen gel matrix culture[J].Investigative Ophthalmology & Visual Science,1993,34(7):2316- 2324.
[190] Griffith M, Osborne R, Xiong X, et al. Functional human corneal equivalents constructed from cell lines[J].Science,1999,286(5447):2169-2172.
[191] 陈家祺,张胜,郭琳洁.体外培养重建角膜组织的技术探讨[J].中国实用眼科杂志,2001, 19(11):815-817.
[192] Lindberg K, Brown M E, Chaves H V, et al. In vitro preparation of human ocular surface epithelial cells for transplantation[J].Investigative Ophthalmology & Visual Science,1993,34:2672-2679.
[193] Kinoshita S. Ocular surface reconstruction by tissue engineering[J].Nippon Ganka Gakkai Zasshi,2002,106(12):837-868.
[194] Ray J T, Li L M, Chen J K. Reconstruction of Damaged Corneas by Transplantation of Autologous Limbal Epithelial Cells[J].New England Journal of Medicine,2000,343:86- 93.
[195] Behrouz A, Harry M. Germ cells frommouse and human embryonic stem cells[J]. Reproduction,2006,132(5):699-707.
[196] Rambhatla L, Chiu C P, Kundu P, et al. Generation of hepatocyte-like cells fromhuman embryonic stem cells[J].Cell Transplant,2003,12(1):1-11.
[197] Maxim A, Vodyanik L, Jack A, et al. Human embryonic stem cell-derived CD34 cells: efficient production in the co-culture with OP9 stromal cells and analysis of lymphohematopoietic potential[J].Blood,2005,105(2):617- 626.
[198] Reubinoff B E, Itsykson P. Neural progenitors from human stem cells[J].Nature Biotechnology,2001,19(12):1134-1140.
[199] Kwon Y D, Oh S K, Kim H S, et al. Cellular manipulation of human embryonic stem cells by TAT-PDX1 protein transduction[J].Molecular Therapy,2005,12(1):28-32.
[200] Ahmad S, Stewart R, Yung S,et al. Differentiation of Human Embryonic Stem Cells into Corneal Epithelial-Like Cells by In Vitro Replication of the Corneal Epithelial Stem Cell Niche[J].Stem Cells,2007,25(5):1145-1155.
[201] 黄冰,王智崇,葛坚,等.利用皮肤干细胞横向分化重建角膜上皮的初步研究[J].中华医学杂志, 2004,84:838-842.
[202] Yu L, Ge J, Wang Z C, et al. Preliminary experimental study of induced Diferentiation of embryonic stem cells into corneal epithelialcells[J].Eye Science,2001,17:138- 143.
[203] Inatomi T, Nakamura T, Koizumi N, et al. Midterm results on ocular surface reconstruction using cultivated autologous oral mucosal epithelial transplantation [J].American Journal of Ophthalmology,2006,141(2):267-275.
[204] Araki-Sasaki K, Ohashi Y, Sasabe T, et al. An SV40-immortalized human corneal epithelial cell line and its characterization[J].Investigative Ophthalmology & Visual Science,1995,36:614-621.
[205] Jin Y, Liu Y, Zhang C, et al. Treatment of rabbit cornea wounds with skin epidermalstem cells[J].Acta Academiae Medicinae Sinicae,2005,27(6):674-677.
[206] 彭亮红,柳林.角膜缘干细胞标志物[J].国际眼科杂志,2005,5(5):1032-1036.
[207] Hayashida Y, Nishida K, Yamato M, et al. Ocular surface reconstruction using autologous rabbit oralmucosalepithelial sheets fabricated ex vivo on a temperature-responsive culture surface[J].Investigative Ophthalmology & Visual Science,2005,46(5):1632-1639.
[208] Hata K, Kagami H, Ueda M, et al. The characteristics of cultured mucosal cell sheetasa material for grafting:comparisonwith cultured epidermalcell sheet[J].Annals of Plastic Surgery,1995,34(5):530-538.
[209] Ueda M, Hata K, Horie K, et al. The potentialofor almucosal cells for cultured epithelium: a prelim inary report[J].Annals of Plastic Surgery,1995,35(5):498-504.
[210] Nakamura T, Endo K, Cooper L J, et al. The successful culture and autologous transplantation of rabbit oralmucosal epithelial cells on amniotic membrane[J]. Investigative Ophthalmology & Visual Science,2003,44(1):106-116.
[211] Nishida K, Yamato M, Hayashida Y, et al. Corneal reconstruction with tissue-engineered cell sheets composed ofautologous oral mucosal epithelium[J].New England Journal of Medicine,2004,351(12):1187-1196.
[212] Insler M S, Lopez J G. Heterologous transplantation verus enhance-ment of human corneal endothelium[J].Cornea,1991,10:136-148.
[213] Peters D M, Dowd N, Brandt C, et al. Human papilloma virus E6/E7 genes can expand the lifespan of human corneal fibroblasts[J].In Vitro Cellular & Developmental Biology-Animal,1996,32(5):279-284.
[214] Reichl S, Bednarz J, Muller-Goymann C C. Human corneal equivalent as cell culture model for in vitro drug permeation studies[J].British Journal of Ophthalmology, 2004,88(4):560-565.
[215] Aboalchamat B, Engelmann K, Bohnke M, et al. Morphological and functional analysis of immortalized human corneal endothelial cells after transplantation[J]. Experimental Eye Research,1999,69(5):547-553.
[216] Joyce N C. Proliferative capacity of the corneal endothelium[J].Progress in Retinal and Eye Research,2003,22:359-389.
[217] 易伟斌,胡竹林,徐岩泽,等.羊膜作为载体培养血管内皮细胞替代自体角膜内皮细胞的研究[J].昆明医学院学报,2006,27:1-4.
[218] Engelmann K, Bednarz J, Valtink M. Prospects for endothelial transplantation[J]. Experimental Eye Research,2004,78:573-578.
[219] 丁勇,徐锦堂,陈建苏.组织工程角膜材料研究进展[J].中国病理生理杂志,2002,18:1557- 1560.
[220] Koizumi N, Inatomi T, Sotozono C, et al. Growth factor mRNAandprotein in preserved human amnioticmembrane[J].Current Eye Research,2000,20:173- 177.
[221] Hao Y, Ma D K, Hwang D G, et al.Identification of antiangiogenic and anti inflammatory proteins in human amniotic membrane[J].Cornea,2000,19:348- 352.
[222] Wang D Y, Hsueh Y J, Yang V C, et al. Propagation and Phenotypic Preservation of RabbitLimbal Epithelial Cells on Amniotic Membrane[J].Investigative Ophthalmology & Visual Science,2003,44:4698-4705.
[223] Espana E M, He H, Kawakita T, et al. Human keratocytes cultured on amniotic membrane stroma preserve morphology and express keratocan [J].Investigative Ophthalmology & Visual Science,2003,44(12):5136- 5141.
[224] He Y G, Alizadeh H, Kinoshita K, et al. Experimental transplantation of cultured human limbal and amniotic epitheial cells onto the corneal surface[J].Cornea,1999,18:570- 579.
[225] Mimura T, Yamagami S, Yokoo S, et al. Cultured human corneal endothelial cell transplantation with a collagen sheet in a rabbit model[J].Investigative Ophthalmology & Visual Science,2004,45:2992-2997.
[226] Ebato B, Friend J, Thoft R A. Comparison of central and peripheral human corneal epithelium in tissue culture[J].Investigative Ophthalmology & Visual Science,1987, 28:1450.
[227] 吴静,徐锦堂,赵松滨.培养角膜上皮移植的实验研究[J].中国实用眼科杂志.2001,19:178- 179.
[228] Engelmann K, Drexler D, B?hnke M. Transplantation of adult human or porcine corneal endothelial cells onto human recipients in vitro.Part I:Cell culturing and transplantation procedure[J].Cornea,1999,18:199-206.
[229] B?hnke M, Eggli P, Engelmann K. Transplantation of cultured adult human or porcine endothelial cells onto human recipients in vitro.Part II: Evaluation in the scanning electron microscope[J].Cornea,1999,18:207-213.
[230] Jumblatt M M, Maurice D M, Schwartz B D. A gelatin membrane substrate for the transplantation of tissue cultured cells[J].Transplantation,1980,29: 498-499.
[231] Mohay J, Lange T M, Soltau J B, et al. Transplantation of corneal endothelial cell using a cell carrier device[J].Cornea,1994,13:173-182.
[232] 郭萍,谢立信,董晓光.人角膜缘干细胞在体外不同载体上培养的实验研究[J].眼科研究, 2001,19(4):304-307.
[233] Hadlock T, Singh S, Vacanti J P, et al. Ocular cell monolayers cultured on biodegradable substrates[J].Tissue Eng,1999,5:187-196.
[234] Griffith M, Hakim M, Shimmura S, et al. Artificial human corneas Scaffolds for transplantation and host regeneration[J].Cornea,2002,21(2):54-61.
[235] Shinazaki J, Aiba M, Goto E, et al. Transplantation of human limbal epithelium cultivated on amniotic membrane for the treatment of severe ocular surface disorders[J].Ophthalmology,2002,109:1285-1290.
[236] 顾国贞,白靓,王芳,等.兔角膜缘干细胞体外分离、培养和生物学鉴定的实验研究[J].现代生物医学进展,2007,7:533-536.
[237] 郭萍,史伟云,李绍伟,等.人角膜缘干细胞体外培养后生物学特性的变化[J].眼科新进展, 2002,22:229-231.
[238] 张晓敏,孙慧敏,赵少贞,等.兔角膜缘干细胞两种培养方法的比较[J].眼科研究,2005,23: 575-578.
[239] Stepp M A, Zhu L. Upregulation of alpha 9 integrin and tenascin during epithelial regeneration after debridement in the cornea[J].J Histochem cytochem,1997,45: 189-201.
[240] Hernandez G E E, Theiss C, Steuhl K P, et al. Expression of p63 in response to phorbol ester in human limbal epithelial cells expanded on intact human amniotic membrane[J]. Investigative Ophthalmology & Visual Science,2003,44:2959-2965.
[241] Matic M, Petrov I N, Chen S, et al. Stem cells of the corneal epithelium lack connexins and metabolite transfer capacity[J].Differentiation,1997,61:251-260.
[242] Harkin DG, Barnard Z, Gillies P, et al. Analysis of p63 and cytokeratin expression in a cultivated limbal autograft used in the treatment of limbal stem cell deficiency [J].Br J Ophthalmol,2004,88:1154-1158.
[243] Djalilian A R, Mahesh S P, Koch C A, et al. Survival of Donor Epithelial Cells after Limbal Stem Cell Transplantation[J].Investigative Ophthalmology and Visual Science, 2005,46:803-807.
[244] Du Y, Chen J, Funderburgh J L, et al. Functional reconstruction of rabbit corneal epithelium by human limbal cells cultur ed on amniotic membrane[J].Mol Vis,2003,9: 635-643.
[245] Kim H, Oh Y. The Effect of Limbal Transplantation Versus Amniotic Membrane Transplantation on Limbal Damaged Rabbit Cornea [J].Investigative Ophthalmology & Visual Science,2003,44:904.
[246] Sangwan V S, Matalia H P, Vemuganti G K, et al. Early Results of Penetrating Keratoplasty After Cultivated Limbal Epithelium Transplantation[J].Arch Ophthalmol, 2005,123(3):334-340.
[247] Yeh L K, Chen W L, Li W, et al. Soluble Lumican Glycoprotein Purified from Human Amniotic Membrane Promotes Corneal Epithelial Wound Healing[J].Investigative Ophthalmology & Visual Science,2005,46(2):479-486.
[248] Li H C, Niederkorn J Y, Neelam S, et al. Immunosuppressive Factors Secreted by Human Amniotic Epithelial Cells[J].Investigative Ophthalmology & Visual Science,2005, 46(3):900-907.
[249] Avila M, Espana M, Moreno C, et al. Reconstruction of ocular surface with heterologous limbal epithelium and amniotic membrane in a rabbit model[J].Cornea,2001,20(4):414- 420.
[250] Grueterich M, Espana E M, Tseng S C. Ex vivo expansion of limbal epithelial stem cells: Amniotic membrane serving as a stem cell niche[J].Surv Ophthalmol,2003,48(6):631- 646.
[251] Nakamura T, Koizumi N, Tsuzuki M, et al. Successful regrafting of cultivated corneal epithelium using amniotic membrane as a carrier in severe ocular surface disease[J]. Cornea,2003,22(1):70-71.
[252] Ramaesh K, Dhillon B. Ex vivo expansion of corneal limbal epithelial/stem cells for corneal surface reconstruction[J].Eur J Ophthalmol,2003,13(6):515-524.
[253] Kubo M,Sonada Y,Muramatsu R,Usui M.Immunogenicity of human amniotic membrane inexperimental xenotransplantation[J].Investigative Ophthalmology & Visual Science, 2001,42:1539-1546.
[254] Barfort P, Maurice D. Electrical potential and fluid transport across the corneal endothelium[J].Experimental Eye Research,1974,19:11-19.
[255] Maurice D M. The location of the fluid pump in the cornea[J].The Journal of Physiology, 1972,221:43-54.
[256] Jun A S, Chakravarti S, Edelhauser H F, et al. Aging changes of mouse corneal endothelium and descemet’s membrane[J].Experimental Eye Research,2006,83:890-896.
[257] Slick W C, Mannagh J and Yuhasz Z. Enzymatic removal and pure culture of rabbit corneal endothelial cells[J].Archives of Ophthalmology,1965,73:229-232.
[258] Engelmann K, Friedl P. Growth of human corneal endothelial cells in a serum- reduced medium[J].Cornea,1995,14:62-70.
[259] Diane A B, Haini Y, Danna L Y,et al. Matrix stimulates the proliferation of human corneal endothelial cells in culture[J].Investigative Ophthalmology & Visual Science,1997,38:1119-1129.
[260] Kazunori M, Jonathan D, Yasuhiro O, et al. Effect of donor age on morphologic variation of cultured human corneal endothelial cells[J].Cornea,2001,20:59-63.
[261] Pan T, Zhao J, Fu Y, et al. Establishment of a novel corneal endothelial cell line from domestic rabbit, Oryctolagus curiculus[J].Science in China Series C:Life Sciences,2007,50:161-169.
[262] Joo C K, Green W R, Pepose J S, et al. Repopulation of denuded murine descemet’s membrane with life-extended murine corneal endothelial cells as a model for corneal cell transplantation[J].Graefe's Archives for Clinical and Experimental Ophthalmology,2000,238:174-180.
[263] Savion N, Isaacs J D, Shuman M A, et al. Proliferation and differentiation of bovine corneal endothelial cells in culture[J].Metabolic Pediatric Systemic Ophthalmology, 1982,6:305-320.
[264] Ogita Y, Nakamura T, Higuchi S, et al. Histochemical studies of mitochondrial activities of cultured corneal endothelial cells of cat during wound-healing[J]. Japanese Journal of Ophthalmology,1990,34:200-215.
[265] Koizumi N, Sakamoto Y, Okumura N, et al. Cultivated corneal endothelial cell sheet transplantation in a primate model [J].Investigative Ophthalmology & Visual Science, 2007,48:4519-4526.
[266] Cintia S P, Zhuo C, Rosa M C, et al. ABCG2 transporter identifies a population of clonogenic human limbal epithelial cells[J].Stem Cell,2005,23:63-73.
[267] Konomi K, Joyce N C. Age and topographical comparison of telomere lengths in human corneal endothelial cells[J].Molecular Vision,2007,13:1251-1258.
[268] Suh L H, Zhang C, Chuck R, et al. Cryopreservation and lentiviral-mediated genetic modification of human primary cultured corneal endothelial cells[J].Investigative Ophthalmology & Visual Science,2007,48:3056-3061.
[269] Stocker F W, Eiring A, Georgiade R, et al. A tissue culturetechnique for growingcorneal epithelial, stromal,and endothelialtissues separately[J].Am J Ophthalmol, 1958,46:294-298.
[270] Zhu C C, Joyce N C. Proliferative response of corneal endothelial cells from young and older donors[J].Investigative Ophthalmology & Visual Science,2004,45:1743- 1751.
[271] Joyce N C, Zhu C C. Human corneal endothelial cell proliferation: potential for use in regenerative medicine[J].Cornea,2004,23:S8-S19.
[272] Woost P G, Jumblatt M M, Eiferman R A, et al. Growth factors and corneal endothelial cells: I. Stimulation of bovine corneal endothelial cell DNA synthesis by defined growth factors[J].Cornea,1992:11:1-10.
[273] Raymond G M, Jumblatt M M, Bartels S P, et al. Rabbit corneal endothelial cells in vitro: effects of EGF[J].Investigative Ophthalmology & Visual Science,1986,27: 474-479.
[274] Nayak S K, Samples J R, Deg J K, et al. Growth characteristics of primate (baboon) corneal endothelium in vitro[J].Investigative Ophthalmology & Visual Science,1986,27:607-611.
[275] Germain L, Auger F A. Tissue engineered biomaterials: biological and mechanical characteristics. Encyclopedic Handbook of Biomaterials and Bioengineering,part B; Applications[M].New York,USA:Marcel Dekker,inc.Publishers,1995,699-734.
[276] Yue BY, Niedra R, Baum J L. Effects of ascorbic acid on cultured rabbit corneal endothelial cells[J].Investigative Ophthalmology & Visual Science,1980,19:1471- 1476.
[277] Wilson S E, Weng J, Blai S, et al. Expression of E6/E7 or SV40 Large T Antigen- Coding Oncogenes in Human Corneal Endothelial Cells Indicates Regulated High-Proliferative Capacity[J].Investigativc Ophthalmology & Visual Science,1995,36:32-40.
[278] Minamino T, Komuro I.Role of telomere in endothelial dysfunction in atherosclerosis[J].Curr Opin Lipidol,2002,13:537-543.
[279] 谢晓繁,贾赤宇,陈璧,等.端粒酶活性对人表皮干细胞分化影响的实验研究[J].实用医学杂志,2006,22:1480-1482.
[280] Murasawa S, Llevadot J, Silver M, et al. Constitutive human telomerase reverse transcriptase expression enhances regenera2 Live properties of endothelial progenitor cells[J].Circulation,2002,106:1133-1139.
[281] Takamasa W, Norikuni S, Westerman K A, et al. Establishment of immortalized human hepatic stellate scavenger cells to develop bioartificial livers[J].Transplantation 2003,75:1873-1880.
[282] Wege H, Le H T, Chui M S, et al. Telomerase reconstition immortalizes human fetal hepatocytes without disrupting their differentiation potential[J].Gastroenterology, 2003,124:432-444.
[283] Turnpenny L, Brickwood S, Spalluto C M, et al. Derivation of Human Embryonic Germ Cells:An alternative source of pluripotent stem cells[J].Stem Cells,2003,21:598- 609.
[284] Bekaert S, De Meyer T, Van O P. Telomere attrition as ageing biomarker[J]. Anticancer Res,2005,25:3011-3021.
[285] Egan C A, Savre-Train I, Shay J W, et al. Analysis of Telomere Lengths in Human Corneal Endothelial Cells from Donors of Different Ages[J].Investigative Ophthalmology & Visual Science,1998,39:648-653.
[286] Whikehart D R, Register S J, Chang Q, et al. Relationship of Telomeres and p53 in Aging Bovine Corneal Endothelial Cell Cultures[J].Investigative Ophthalmology & Visual Science,2000,41:1070-1075.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |