BMP9对心肌前体干细胞改善心梗大鼠心功能的影响
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摘要
第一部分AdBMP9和AdGFP的扩增及心肌前体细胞的感染
目的:通过293细胞扩增得到高滴度的腺病毒,用AdBMP9重组腺病毒感染心肌前体干细胞使之高表达BMP9。
方法:培养并利用293细胞对低滴度的AdBMP9和AdGFP腺病毒进行反复扩增后收集得到高滴度腺病毒。当心肌前体干细胞传代贴壁至70%~80%时,在5 ml完全培养基中加入20μL扩增后的腺病毒感染心肌前体干细胞6～8小时后换液,24小时后荧光倒置显微镜观察绿色荧光表达并用流式细胞仪检测细胞感染效率,RT-PCR技术检测被感染的心肌前体干细胞BMP9的表达量。
结果:
1.病毒原液经293细胞反复扩增后,获得了高滴度的AdBMP9和AdGFP。
2.高滴度的AdBMP9和AdGFP分别感染心肌前体干细胞24小时后,流式细胞仪检测荧光表达效率分别为46%和45%。
3.RT-PCR结果显示AdBMP9感染的心肌前体干细胞BMP9的表达量明显增高(P<0.05),而AdGFP感染的心肌前体干细胞中BMP9的表达量跟心肌前体干细胞本身相比并无明显的差异(P>0.05)。
结论:利用293细胞扩增可以得到高滴度的腺病毒;高滴度的腺病毒对心肌前体干细胞有较高的感染率;AdBMP9感染心肌前体干细胞后BMP9的表达量明显增高。
第二部分BMP9对心肌前体干细胞改善心梗大鼠心功能作用的研究
目的:探讨BMP9能否促进心肌前体干细胞改善心梗大鼠心功能。
方法:26只雄性SD大鼠随机分成三组(1)MI组(n=10);(2)MI+GFP组(n=8);(3)MI+BMP9组(n=8)。动脉圆锥下结扎冠状动脉左前降支10~15min后,在梗死心肌组织周围即时多点注入生理盐水混匀的腺病毒感染的心肌前体干细胞悬液(100μL,5～6×106个/mL)。细胞植入四周后,利用超声心动图检测左室心功能参数,Masson染色计算心脏梗死面积及观察植入细胞形态学和免疫荧光检测α-MHC及α-actin蛋白的表达。
结果:
1.动脉圆锥下结扎大鼠冠状动脉的左前降支后,成功建立心梗模型。术后4周形态学显示,心肌梗死部位心肌细胞稀少,胞浆稀薄,有大量的纤维组织及炎症细胞,且左心室壁明显变薄。
2.超声心动图结果显示细胞植入4周时,MI+BMP9组LVESD较MI组降低而FS则升高,均有显著性差异(P﹤0.05 , P﹤0.01);且MI+ BMP9组与MI+GFP组比较,改善更为明显(P﹤0.05)。
3.MI组大鼠心肌梗死面积的百分比为39.7%±2%(n=8),MI+GFP组的心梗面积百分比为37.3%±3%(n=6),MI+BMP9组的心梗面积百分比为34.6%±3%(n=7)。MI+GFP组同MI组比较(P﹤0.05),MI+BMP9组同MI组比较(P﹤0.01)且MI+BMP9组同MI+GFP组相比P﹤0.05)。
4.植入细胞4周后,形态学检查发现植入的细胞离正常心肌组织越接近,细胞胞浆越饱满,而在注入细胞团块中央的细胞胞浆稀少。免疫荧光结果显示,高表达BMP9的心肌前体干细胞和GFP标记的心肌前体细胞均有a-MHC和a-actin的表达。通过测量分析各组目的蛋白荧光的吸光度值发现,MI+BMP9组中植入细胞的a-MHC和a-actin表达量较周围的正常组织弱(P﹤0.05),但较MI+GFP组中植入细胞的表达较高(P﹤0.05)。
结论: BMP9可以促进心肌前体干细胞分化表达心肌特异性结构蛋白,并增强心梗大鼠的左室心功能。
PART ONE : AMPLIFICATION OF AdBMP9 AND AdGFP, THE INFECTION OF CARDIAC PROGENTIOR CELLS
Objective: Amplifying the AdGFP and AdBMP9 with 293 cells to get high-titer adenovirus; Infect the CPCs with AdBMP9 and improve the expression of BMP9 of the CPCs.
Methord: Cultivate the 293 cells and amplify the adenovirus with 293 cells. Cultivate the cardiac progenitor cells. CPCs were infected by AdBMP9 which express green fluorescent protein. After twenty-four hours, the expression of BMP9 in CPCs were tested by semi-quantitative RT-PCR.
Result:
1. We got high titer adenovirus by amplifying the AdGFP and AdBMP9 with 293 cells.
2. After CPCs infected with adenovirus 24 hours, the efficiency of AdGFP and AdBMP9 is 45% and 46%.
3. In three groups, the expression of BMP9 is improved dramatically in CPCs infected with AdBMP9.
Conclusion : The titer of AdBMP9 and AdGFP is improved though 293 amplification. The expression of BMP9 is improved dramatically in CPCs infected with AdBMP9.
PART TWO: THE STUDY OF BMP9 IN CPCs AMELIORATE HEART FUNCTION IN THE RAT MODEL OF MYOCARDIAL INFARCTION
Objective: To test whether cardiac progenitor cells (CPCs) which induced by bone morphogenetic proteins9 (BMP9) could improve the heart function or not.
Methord: Twenty six male rats were divided into three groups. MI group (n=10) ,MI+GFP group (n=8), MI+BMP9 group (n=8.). Four weeks after transplantion, the change of CPCs morphology was tested though the expression of cardiac-specific structural proteins alpha myosin heavy chain(a-MHC) and a-actin of the implanted CPCs with HE staining and immunofluorescent analysis. The myocardium infarct area was detected though Masson staining. Heart function was evaluated by echocardiography, in preoperative and postoperative four weeks.
Result:
1.MI models were established successfully by blocking left anterior descending coronary arteries.
2. Four weeks after transplantation , the heart function of the rats were compared during the three groups. In LVESD and FS level , MI+BMP9 group better than other group(p<0.05). The level of LVESD and FS in MI+GFP group was improved compared with the MI group.
3. Four weeks after transplantation, the implanted CPCs reduced the size of infracted obviously(MI+GFP VS MI ,P﹤0.05)and could be promoted by BMP9(MI+BMP9 VS MI P﹤0.01)
4. The result of HE staining and immunofluorescent showed that the grafted CPCs could fuse with the normal cardiomyocytes and expressed the cardiac-specific proteins a-MHC and a-actin, at four weeks after transplantation. But the expression of cardiac-specific structural proteins is lower than the abnormal cardiac tissue. The expression of a-MHC and a-actin in MI+BMP9 group is more than MI+GFP group.( P﹤0.05) Conclusion:BMP9 could improve the expression of cardiac-specific structural proteins of CPCs and promote cardiac progenitor cells ameliorate the heart function in the rat model of myocardial infarction.
目的:通过293细胞扩增得到高滴度的腺病毒,用AdBMP9重组腺病毒感染心肌前体干细胞使之高表达BMP9。
方法:培养并利用293细胞对低滴度的AdBMP9和AdGFP腺病毒进行反复扩增后收集得到高滴度腺病毒。当心肌前体干细胞传代贴壁至70%~80%时,在5 ml完全培养基中加入20μL扩增后的腺病毒感染心肌前体干细胞6～8小时后换液,24小时后荧光倒置显微镜观察绿色荧光表达并用流式细胞仪检测细胞感染效率,RT-PCR技术检测被感染的心肌前体干细胞BMP9的表达量。
结果:
1.病毒原液经293细胞反复扩增后,获得了高滴度的AdBMP9和AdGFP。
2.高滴度的AdBMP9和AdGFP分别感染心肌前体干细胞24小时后,流式细胞仪检测荧光表达效率分别为46%和45%。
3.RT-PCR结果显示AdBMP9感染的心肌前体干细胞BMP9的表达量明显增高(P<0.05),而AdGFP感染的心肌前体干细胞中BMP9的表达量跟心肌前体干细胞本身相比并无明显的差异(P>0.05)。
结论:利用293细胞扩增可以得到高滴度的腺病毒;高滴度的腺病毒对心肌前体干细胞有较高的感染率;AdBMP9感染心肌前体干细胞后BMP9的表达量明显增高。
第二部分BMP9对心肌前体干细胞改善心梗大鼠心功能作用的研究
目的:探讨BMP9能否促进心肌前体干细胞改善心梗大鼠心功能。
方法:26只雄性SD大鼠随机分成三组(1)MI组(n=10);(2)MI+GFP组(n=8);(3)MI+BMP9组(n=8)。动脉圆锥下结扎冠状动脉左前降支10~15min后,在梗死心肌组织周围即时多点注入生理盐水混匀的腺病毒感染的心肌前体干细胞悬液(100μL,5～6×106个/mL)。细胞植入四周后,利用超声心动图检测左室心功能参数,Masson染色计算心脏梗死面积及观察植入细胞形态学和免疫荧光检测α-MHC及α-actin蛋白的表达。
结果:
1.动脉圆锥下结扎大鼠冠状动脉的左前降支后,成功建立心梗模型。术后4周形态学显示,心肌梗死部位心肌细胞稀少,胞浆稀薄,有大量的纤维组织及炎症细胞,且左心室壁明显变薄。
2.超声心动图结果显示细胞植入4周时,MI+BMP9组LVESD较MI组降低而FS则升高,均有显著性差异(P﹤0.05 , P﹤0.01);且MI+ BMP9组与MI+GFP组比较,改善更为明显(P﹤0.05)。
3.MI组大鼠心肌梗死面积的百分比为39.7%±2%(n=8),MI+GFP组的心梗面积百分比为37.3%±3%(n=6),MI+BMP9组的心梗面积百分比为34.6%±3%(n=7)。MI+GFP组同MI组比较(P﹤0.05),MI+BMP9组同MI组比较(P﹤0.01)且MI+BMP9组同MI+GFP组相比P﹤0.05)。
4.植入细胞4周后,形态学检查发现植入的细胞离正常心肌组织越接近,细胞胞浆越饱满,而在注入细胞团块中央的细胞胞浆稀少。免疫荧光结果显示,高表达BMP9的心肌前体干细胞和GFP标记的心肌前体细胞均有a-MHC和a-actin的表达。通过测量分析各组目的蛋白荧光的吸光度值发现,MI+BMP9组中植入细胞的a-MHC和a-actin表达量较周围的正常组织弱(P﹤0.05),但较MI+GFP组中植入细胞的表达较高(P﹤0.05)。
结论: BMP9可以促进心肌前体干细胞分化表达心肌特异性结构蛋白,并增强心梗大鼠的左室心功能。
PART ONE : AMPLIFICATION OF AdBMP9 AND AdGFP, THE INFECTION OF CARDIAC PROGENTIOR CELLS
Objective: Amplifying the AdGFP and AdBMP9 with 293 cells to get high-titer adenovirus; Infect the CPCs with AdBMP9 and improve the expression of BMP9 of the CPCs.
Methord: Cultivate the 293 cells and amplify the adenovirus with 293 cells. Cultivate the cardiac progenitor cells. CPCs were infected by AdBMP9 which express green fluorescent protein. After twenty-four hours, the expression of BMP9 in CPCs were tested by semi-quantitative RT-PCR.
Result:
1. We got high titer adenovirus by amplifying the AdGFP and AdBMP9 with 293 cells.
2. After CPCs infected with adenovirus 24 hours, the efficiency of AdGFP and AdBMP9 is 45% and 46%.
3. In three groups, the expression of BMP9 is improved dramatically in CPCs infected with AdBMP9.
Conclusion : The titer of AdBMP9 and AdGFP is improved though 293 amplification. The expression of BMP9 is improved dramatically in CPCs infected with AdBMP9.
PART TWO: THE STUDY OF BMP9 IN CPCs AMELIORATE HEART FUNCTION IN THE RAT MODEL OF MYOCARDIAL INFARCTION
Objective: To test whether cardiac progenitor cells (CPCs) which induced by bone morphogenetic proteins9 (BMP9) could improve the heart function or not.
Methord: Twenty six male rats were divided into three groups. MI group (n=10) ,MI+GFP group (n=8), MI+BMP9 group (n=8.). Four weeks after transplantion, the change of CPCs morphology was tested though the expression of cardiac-specific structural proteins alpha myosin heavy chain(a-MHC) and a-actin of the implanted CPCs with HE staining and immunofluorescent analysis. The myocardium infarct area was detected though Masson staining. Heart function was evaluated by echocardiography, in preoperative and postoperative four weeks.
Result:
1.MI models were established successfully by blocking left anterior descending coronary arteries.
2. Four weeks after transplantation , the heart function of the rats were compared during the three groups. In LVESD and FS level , MI+BMP9 group better than other group(p<0.05). The level of LVESD and FS in MI+GFP group was improved compared with the MI group.
3. Four weeks after transplantation, the implanted CPCs reduced the size of infracted obviously(MI+GFP VS MI ,P﹤0.05)and could be promoted by BMP9(MI+BMP9 VS MI P﹤0.01)
4. The result of HE staining and immunofluorescent showed that the grafted CPCs could fuse with the normal cardiomyocytes and expressed the cardiac-specific proteins a-MHC and a-actin, at four weeks after transplantation. But the expression of cardiac-specific structural proteins is lower than the abnormal cardiac tissue. The expression of a-MHC and a-actin in MI+BMP9 group is more than MI+GFP group.( P﹤0.05) Conclusion:BMP9 could improve the expression of cardiac-specific structural proteins of CPCs and promote cardiac progenitor cells ameliorate the heart function in the rat model of myocardial infarction.
引文
[1]. Orlic D, Kajstura J,et al.Bone marrow cells regenerate infarcted myocardium. [J] Nature 2001;410:701-705.
[2]. Strauer BE,Brehm M, et al. Repair of infrcted myocardium by autologous intracoronary mononuclear bone marrow cell transplantation in human. [J] Circulation 2002; 106:1913-1918.
[3]. Keiichi Fukuda, Shinsuke Yuasa. Stem Cells as a Source of Regenerative Cardiomyocytes; [J]Circ. Res. 2006 Apr 28;98(8):1002-13
[4]. Kenneth R. Boheler, Jaroslaw Czyz, David Tweedie, et al. Differentiation of Pluripotent Embryonic Stem Cells Into Cardiomyocytes; [J]Circ. Res. 2002 Aug 9;91(3):189-201;
[5]. Kawai T, Takahashi T ,Esaki M, et al. Efficient Cardiomyogenic Differentiation of Embryonic Stem Cell by Fibroblast Growth Factor 2 and Bone Morphogenetic Protein 2. [J]Circ J. 2004 Jul;68(7):691-702;
[6]. ]Li TS, Komota T, Ohshima M,et al .TGF-b induces the differentiation of bone marrow stem cells into immature cardiomyocytes;[J]Biochem Biophy Res Commun.2008 Feb 22;366(4):1074-80;
[7]. Susana M. Chuva De Sousa Lopes , Alie Feijen , Jeroen Korving, et al. ConnectiveTissue Growth Factor Expression and Smad Signaling During Mouse Heart Development and Myocardial Infarction; [J] Dev Dyn .2004 Nov;231(3):542-50;
[8]. Laura Lagostenaa, Daniele Avitabileb, Elena De Falcoa ,et al. Electrophysiological properties of mouse bone marrow c-kit+ cells co-cultured onto neonatal cardiac myocytes;[J]Cardiovasc Res. 2005 Jun 1;66(3):482-92;
[9] Chang Sung-A, Lee EJ ,Kang HJ, et al. Impact of Myocardial Infarct Proteins and Oscillating Pressure on the Differentiation of Mesenchymal Stem Cells: Effect of Acute Myocardial Infarction on Stem Cell Differentiation;[J]Stem Cells .2008 Jul;26(7):1901-12.
[10] Rajasingh J, Bord E, Hamada H, et al. STAT3-Dependent Mouse Embryonic Stem Cell Differentiation Into Cardiomyocytes Analysis of Molecular Signaling and Therapeutic Efficacy of Cardiomyocyte Precommitted mES Transplantation in a Mouse Model of Myocardial Infarction;[J]Circ. Res. 2007 Oct 26;101(9):910-8.
[11] Lim JY, Kim WH, Kim J, et al. Involvement of TGF-β1 Signaling in Cardiomyocyte Differentiation from P19CL6 Cells; [J]Mol. Cells, 2007 Dec 31;24(3):431-6
[12] Choi SC, Yoon J, Shim WJ, et al. 5-azacytidine induces cardiac differentiation of P19 embryonic stem cells; [J] Exp Mol Med, 2004 Dec 31;36(6):515-23.
[13] Boudewijn P.T. Kruithof, Bram van Wijk, Semir Somi,et al. BMP and FGF regulate the differentiation of multipotential pericardial mesoderm into the myocardial or epicardial lineage; [J] Dev Biol. 2006 Jul 15;295(2):507-22.
[14] Singla DK, Sobel BE. Enhancement by growth factors of cardiac myocyte differentiation from embryonic stem cells: A promising foundation for cardiac regeneration; [J] Biochem and Biophys Res Commun. 2005 Sep 30;335(3):637-42
[15] Jiwang Zhanga, Linheng Li. BMP signaling and stem cell regulation. Developmental Biology [J]2005,284 ,1– 11
[16] Coviella CI, Follettie MT, Mellott TJ, et al.Bone morphogenetic protein 9 induces the transcriptome of basal forebrain cholinergic neurons. [J] Proc Natl Acad Sci U S A ,2005 ,102(19):6984-9.
[17] David L, LamandéN, Gasc JM, et al.Bone morphogenetic protein-9 is a circulating vascular quiescence factor. [J] Circ Res. 2008,102(8): 914-22.
[18] Tang N, Song WX, Luo J, et al.BMP9-induced osteogenic differentiation of mesenchymal progenitors requires functional canonical Wnt/beta-catenin signaling .[J] Cell Mol Med. 2008,[Epub ahead of print]
[1]. Baltzer Aw, Lattermann C, et al. Genetic enhancement of fracture repair: healing of experimental segmental defect by adenoviral transfer of the BMP2 gene[J].Gene Ther , 2000, 7(9): 734-739.
[2]. Zhao J, Pettigrew GJ, et al. Lentiviral vectors for delivery of genes into neonatal and adult ventricular cardiac myocytes in vitro and in vivo. [J] Basic Res Cardiol 2002;97:348-358
[3]. Mizuguchi H, Koizumi N,et.al. CAR- or alphav integrin-binding ablated adenovirus vectors, but not fiber modified vectors containing RGD peptide, do not change the systemic gene transfer properties in mice. [J] Gene Ther 2002;9:769-776
[4]. Havenga MJ, Lemckert AA,et al. Exploiting the natural diversity in adenovirus tropism for therapy and prevention of disease. [J] J Virol 2002;76:4612-4620
[5]. Wang G, Berk AJ. In Vivo association of adenovirus large E1A protein with the human mediator complex in adenovirusinfected and -Transformed cells. [J] J Virol 2002;76:9186-9193
[6]. Stevens JL, Cantin GT, et al. Transcription control by E1A and MAP kinase pathway via Sur2 mediator subunit. [J] Science 2002;296: 755-758
[7]. Nada S, Trempe JP. Characterization of adeno-associated virus rep protein inhibition of adenovirus E2a gene expression. [J]Virology 2002;293:345-355
[8]. Jing XJ, Kalman-Maltese V,et.al. Inhibition of adenovirus cytotoxicity, replication, and E2a gene expression by adeno-associated virus. [J] Virology 2001;91:140-151
[9]. Pandori M, Hobson D, et.al Adenovirus-microbead conjugatespossess enhanced infectivity: a new strategy for localizedgene delivery. [J]Virology 2002;299: 204-212
[10]. Nasz I, Adam E. Recombinant adenovirus vectors for genetherapy and clinical trials. [J] Acta Microbiol Immunol Hung 2001;48:323-348
[11]. Adam E, Nasz I. Adenovirus vectors and their clinical application in gene therapy. [J]Orv Hetil 2001;142:2061-2070.
[12].赫春秋,冯志华等人,腺病毒载体的特点及其在HCV研究的应用[J].世界胃肠杂志.2003 ;11(6):803-805
[13]. Beltrami AP, Barlucchi L, Torella D,et al .Adult Cardiac Stem Cells Are Multipotent and Support Myocardial Regeneration[J]. Cell, 2003, 114(6):763 -76.
[14]. Coviella CI, Follettie MT, Mellott TJ, et al.Bone morphogenetic protein 9 induces the transcriptome of basal forebrain cholinergic neurons[J]. Proc Natl Acad Sci U S A ,2005 ,102(19):6984-9.
[15]. David L, LamandéN, Gasc JM, et al.Bone morphogenetic protein-9 is a circulating vascular quiescence factor[J].Circ Res. 2008,102(8): 914-22.
[16]. Tang N, Song WX, Luo J, et al.BMP9-induced osteogenic differentiation of mesenchymal progenitors requires functional canonical Wnt/beta-catenin signaling[J].J Cell Mol Med. 2008,[Epub ahead of print]
[1]. Vulliet YS, Greeley M. et al.Intra-coronary arterial injection of mesenchymal stromal cells and microinfarction in dogs. [J] Lancet.2004; 362(9411) :783-784;
[2]. Yoon YS ,Park JS,et al. Unexpected severe calcification after transplantation of bone marrow cells in acute myocardial infaration. [J] Circulation. 2004; 109(25): 3254-3157
[3]. Houghton JM, Stoicov C,et al. Gastric ancer originating from bone marrow derived cells. [J]Science.2004;306(5071):1568-1571
[4]. Yang lei,Mark,H et.al Human cardiovascular progenitor cells develop from a KDR+ embryonic-stem-cell-derived population;[J],Nature 2008 453, 524-528
[5]. Mazhari R Hare JM. Advances in cell-based therapy for structural heart disease;[J] Prog Cardiovasc Dis. 2007 May-Jun;49(6):387-95.
[6].向平,宋银子,陈沅等,提高大鼠心肌梗死模型成功率及存活率的研究。[J]第三军医大学学报,2008;30:4,329-332。
[7]. Ohara F,Sugimoto T, et al. Protective Effect of FR168888, a New Na+/H+ Exchange Inhibitor, on Ischemia and Reperfusion-Induced Arrhythmia and Myocardial infarction in Rats:In Comarison With Other Cardioprotetive Compounds. [J] Jpn.J. Pharmacol, 1999;80:295-302.
[8]. Tousoulis D, Briasoulis A. et.al. Heart regeneration:;what cells to use and how? [J]Curr Opin Pharmacol. 2008 Apr;8(2):211-8. Epub 2008 Mar 4.;
[9]. Zeng Bin, Lin-Guo Sheng. et.al Efficient cardiomyocyte differentiation of embryonic stem cells by bone morphogenetic protein-2 combined with visceralendoderm-like cells.[J] Cell Biology International 2006, 769-776;
[10]. Rebecca Y. Kim, Elizabeth J. Robertson, et al. Bmp6 and Bmp7 Are Required for Cushion Formation and Septation in the Developing Mouse Heart;[J] Developmental Biology ,2001,235, 449–466
[11]. Taha MF, Valojerdi MR,et.al. Effect of bone morphogenetic protein-4 (BMP-4) on cardiomyocyte differentiation from mouse embryonic stem cell; [J] Int J Cardiol. 2007, Aug 9;120(1):92-101.
[12]. Coviella CI, Follettie MT, Mellott TJ, et al.Bone morphogenetic protein 9 induces the transcriptome of basal forebrain cholinergic neurons. [J] Proc Natl Acad Sci U S A ,2005 ,102(19):6984-9.
[13]. David L, LamandéN, Gasc JM, et al.Bone morphogenetic protein-9 is a circulating vascular quiescence factor. [J]Circ Res. 2008,102(8): 914-22.
[14]. Tang N, Song WX, Luo J, et al.BMP9-induced osteogenic differentiation of mesenchymal progenitors requires functional canonical Wnt/beta-catenin signaling. [J] Cell Mol Med. 2008,[Epub ahead of print]
[15]. Buddhadeb Dawn, Adam B. Stein , et al. Cardiac stem cells delivered intravascularly traverse the vessel barrier, regenerate infarcted myocardium, and improve cardiac function;[J] Proc Natl Acad Sci USA, 2005, 102(10):3766-3771
[16].黄枫,余志斌等.心肌肌原纤维的分子结构与功能.[J]心脏杂志;2000,12,(1);
[17].谭金童,陈沅等.BMP9对C3H10T1/2干细胞向心肌样细胞分化的影响[J].重庆医科大学学报,2009,Vol.34 NO.6
[1]. Keiichi Fukuda, Shinsuke Yuasa. Stem Cells as a Source of Regenerative Cardiomyocytes; [J] Circ. Res. 2006 Apr 28;98(8):1002-13.
[2]. Kenneth R. Boheler, Jaroslaw Czyz, David Tweedie, et al. Differentiation of Pluripotent Embryonic Stem Cells Into Cardiomyocytes; [J] Circ. Res. 2002 Aug 9;91(3):189-201
[3]. Kawai T, Takahashi T ,Esaki M, et al. Efficient Cardiomyogenic Differentiation of Embryonic Stem Cell by Fibroblast Growth Factor 2 and Bone Morphogenetic Protein 2;[J]Circ J. 2004 Jul;68(7):691-702
[4]. Li TS, Komota T, Ohshima M,et al .TGF-b induces the differentiation of bone marrow stem cells into immature cardiomyocytes;[J]Biochem Biophy Res Commun.2008 Feb 22;366(4):1074-80
[5]. Susana M. Chuva De Sousa Lopes , Alie Feijen , Jeroen Korving, et al. Connective Tissue Growth Factor Expression and Smad Signaling During Mouse HeartDevelopment and Myocardial Infarction; [J]Dev Dyn .2004 Nov;231(3):542-50
[6]. Laura Lagostenaa, Daniele Avitabileb, Elena De Falcoa ,et al. Electrophysiological properties of mouse bone marrow c-kit+ cells co-cultured onto neonatal cardiac myocytes;[J]Cardiovasc Res. 2005 Jun 1;66(3):482-92。
[7]. Chang Sung-A, Lee EJ ,Kang HJ, et al. Impact of Myocardial Infarct Proteins and Oscillating Pressure on the Differentiation of Mesenchymal Stem Cells: Effect of Acute Myocardial Infarction on Stem Cell Differentiation;[J]Stem Cells .2008 Jul;26(7):1901-12.
[8]. Rajasingh J, Bord E, Hamada H, et al. STAT3-Dependent Mouse Embryonic Stem Cell Differentiation Into Cardiomyocytes Analysis of Molecular Signaling and Therapeutic Efficacy of Cardiomyocyte Precommitted mES Transplantation in a Mouse Model of Myocardial Infarction;[J]Circ. Res. 2007 Oct 26;101(9):910-8.
[9]. Lim JY, Kim WH, Kim J, et al. Involvement of TGF-β1 Signaling in Cardiomyocyte Differentiation from P19CL6 Cells; [J]Mol. Cells, 2007 Dec 31;24(3):431-6
[10]. Choi SC, Yoon J, Shim WJ, et al. 5-azacytidine induces cardiac differentiation of P19 embryonic stem cells; [J]Exp Mol Med, 2004 Dec 31;36(6):515-23.
[11]. Boudewijn P.T. Kruithof, Bram van Wijk, Semir Somi,et al. BMP and FGF regulate the differentiation of multipotential pericardial mesoderm into the myocardial or epicardial lineage; [J] Dev Biol. 2006 Jul 15;295(2):507-22.
[12]. Singla DK, Sobel BE. Enhancement by growth factors of cardiac myocyte differentiation from embryonic stem cells: A promising foundation for cardiac regeneration; [J]Biochem and Biophys Res Commun. 2005 Sep 30;335(3):637-42
[13]. Van Wijk B, Moorman AF, van den Hoff MJ. Role of bone morphogenetic proteins in cardiac differentiation; [J]Cardiovasc Res. 2007 May 1;74(2):244-55.
[14]. Sabine Mazerbourg, Katrin Sangkuhl, Ching-Wei Luo, et al. Identification of Receptors and Signaling Pathways for Orphan Bone Morphogenetic Protein/Growth Differentiation Factor Ligands Based on Genomic Analyses; [J] J Bio Chem. 2005 Sep 16;280(37):32122-32.
[15]. Zhang J, Li L .BMP signaling and stem cell regulation;[J]Devl Bio. 2005 Aug 1;284(1):1-11
[16]. Zeng YA, Rahnama M, Wang S, et al.Drosophila Nemo antagonizes BMP signalingby phosphorylation of Mad and inhibition of its nuclear accumulation ;[J]Development 134, 2061-2071 (2007);
[17]. Shiojioma I, Walsh K. Regulation of cardiac growth and coronary angiogenesis by the Akt/PKB signaling pathway;[J]Genes Dev. 2006 Dec 15;20(24):3347-65 .
[18]. Klaus A, Saga Y, Taketo MM, et al.Distinct roles of Wnt/_-catenin and Bmp signaling during early cardiogenesis;[J]Proc Natl Acad Sci U S A. 2007 Nov 20;104(47):18531-6.
[19]. Papadimou E, Ménard C, Grey C, et al. Interplay between the retinoblastoma protein and LEK1 specifies stem cells toward the cardiac lineage; [J]EMBO J. 2005 May 4;24(9):1750-61.
[20]. Balemans W, Van Hul W. Extracellular regulation of BMP signaling in vertebrates: a cocktail of modulators.; [J]Dev Bio. 2002 Oct 15;250(2):231-50.
[21]. Doss MX, Chen S, Winkler J, et al. Transcriptomic and phenotypic analysis of murine embryonic stem cell der ived BMP2+ lineage cells: an insight into mesodermal patterning; [J] Genome Biol.2007;8(9):R184.
[22]. Taha MF, Valojerdi MR, Mowla SJ. Effect of bone morphogenetic protein-4 (BMP-4) on cardiomyocyte differentiation from mouse embryonic stem cell; [J]Int J Cardiol. 2007 Aug 9;120(1):92-101.
[23]. Dudley AT, Robertson EJ. Overlapping expression domains of bone morphogenetic protein family members potentially account for limited tissue defects in BMP7 deficient embryos; [J]Dev Dyn. 1997 Mar;208(3):349-62.
[24]. Kim RY, Robertson EJ, Solloway MJ. Bmp6 and Bmp7 are required for cushion formation and septation in the developing mouse heart; [J]Dev Biol. 2001 Jul 15;235(2):449-66.
[25]. Somi S, Buffing AA, Moorman AF, et al. Dynamic patterns of expression of BMP isoforms 2,4,5,6, and 7 during chicken heart development. [J]Anat Rec A Discov Mol cell Evol Bio. 2004 Jul;279(1):636-51 .
[26]. Zhang H, Bradley A. Mice deficient for BMP2 are nonviable and have defects in amnion/chorion and cardiac development. [J]Development. 1996 Oct ; 122 (10) :2977-86.
[27]. Winnier G, Blessing M, Labosky PA, et al. Bone morphogenetic protein-4 is required for mesoderm formation and patterning in the mouse. [J]Gene Dev. 1995Sep 1;9(17):2105-16.
[28]. Kingsley DM, Bland AE, Grubber JM, et al. The mouse short ear skeletal morphogenesis locus is associated with defects in a bone morphogenetic member of the TGF beta superfamily. [J]Cell. 1992 Oct 30;71(3):399-410.
[29]. Solloway MJ, Dudley AT, Bikoff EK, Lyons KM, et al. Mice lacking Bmp6 function. [J] Dev Genet. 1998;22(4):321-39.
[30]. Hahn JY, Cho HJ, Kang HJ, et al. Pre-Treatment of Mesenchymal Stem Cells With a Combination of Growth Factors Enhances Gap Junction Formation, Cytoprotective Effect on Cardiomyocytes, and Therapeutic Efficacy for Myocardial Infarction;[J]J Am Coll Cardiol. 2008 Mar 4;51(9):933-43.
[2]. Strauer BE,Brehm M, et al. Repair of infrcted myocardium by autologous intracoronary mononuclear bone marrow cell transplantation in human. [J] Circulation 2002; 106:1913-1918.
[3]. Keiichi Fukuda, Shinsuke Yuasa. Stem Cells as a Source of Regenerative Cardiomyocytes; [J]Circ. Res. 2006 Apr 28;98(8):1002-13
[4]. Kenneth R. Boheler, Jaroslaw Czyz, David Tweedie, et al. Differentiation of Pluripotent Embryonic Stem Cells Into Cardiomyocytes; [J]Circ. Res. 2002 Aug 9;91(3):189-201;
[5]. Kawai T, Takahashi T ,Esaki M, et al. Efficient Cardiomyogenic Differentiation of Embryonic Stem Cell by Fibroblast Growth Factor 2 and Bone Morphogenetic Protein 2. [J]Circ J. 2004 Jul;68(7):691-702;
[6]. ]Li TS, Komota T, Ohshima M,et al .TGF-b induces the differentiation of bone marrow stem cells into immature cardiomyocytes;[J]Biochem Biophy Res Commun.2008 Feb 22;366(4):1074-80;
[7]. Susana M. Chuva De Sousa Lopes , Alie Feijen , Jeroen Korving, et al. ConnectiveTissue Growth Factor Expression and Smad Signaling During Mouse Heart Development and Myocardial Infarction; [J] Dev Dyn .2004 Nov;231(3):542-50;
[8]. Laura Lagostenaa, Daniele Avitabileb, Elena De Falcoa ,et al. Electrophysiological properties of mouse bone marrow c-kit+ cells co-cultured onto neonatal cardiac myocytes;[J]Cardiovasc Res. 2005 Jun 1;66(3):482-92;
[9] Chang Sung-A, Lee EJ ,Kang HJ, et al. Impact of Myocardial Infarct Proteins and Oscillating Pressure on the Differentiation of Mesenchymal Stem Cells: Effect of Acute Myocardial Infarction on Stem Cell Differentiation;[J]Stem Cells .2008 Jul;26(7):1901-12.
[10] Rajasingh J, Bord E, Hamada H, et al. STAT3-Dependent Mouse Embryonic Stem Cell Differentiation Into Cardiomyocytes Analysis of Molecular Signaling and Therapeutic Efficacy of Cardiomyocyte Precommitted mES Transplantation in a Mouse Model of Myocardial Infarction;[J]Circ. Res. 2007 Oct 26;101(9):910-8.
[11] Lim JY, Kim WH, Kim J, et al. Involvement of TGF-β1 Signaling in Cardiomyocyte Differentiation from P19CL6 Cells; [J]Mol. Cells, 2007 Dec 31;24(3):431-6
[12] Choi SC, Yoon J, Shim WJ, et al. 5-azacytidine induces cardiac differentiation of P19 embryonic stem cells; [J] Exp Mol Med, 2004 Dec 31;36(6):515-23.
[13] Boudewijn P.T. Kruithof, Bram van Wijk, Semir Somi,et al. BMP and FGF regulate the differentiation of multipotential pericardial mesoderm into the myocardial or epicardial lineage; [J] Dev Biol. 2006 Jul 15;295(2):507-22.
[14] Singla DK, Sobel BE. Enhancement by growth factors of cardiac myocyte differentiation from embryonic stem cells: A promising foundation for cardiac regeneration; [J] Biochem and Biophys Res Commun. 2005 Sep 30;335(3):637-42
[15] Jiwang Zhanga, Linheng Li. BMP signaling and stem cell regulation. Developmental Biology [J]2005,284 ,1– 11
[16] Coviella CI, Follettie MT, Mellott TJ, et al.Bone morphogenetic protein 9 induces the transcriptome of basal forebrain cholinergic neurons. [J] Proc Natl Acad Sci U S A ,2005 ,102(19):6984-9.
[17] David L, LamandéN, Gasc JM, et al.Bone morphogenetic protein-9 is a circulating vascular quiescence factor. [J] Circ Res. 2008,102(8): 914-22.
[18] Tang N, Song WX, Luo J, et al.BMP9-induced osteogenic differentiation of mesenchymal progenitors requires functional canonical Wnt/beta-catenin signaling .[J] Cell Mol Med. 2008,[Epub ahead of print]
[1]. Baltzer Aw, Lattermann C, et al. Genetic enhancement of fracture repair: healing of experimental segmental defect by adenoviral transfer of the BMP2 gene[J].Gene Ther , 2000, 7(9): 734-739.
[2]. Zhao J, Pettigrew GJ, et al. Lentiviral vectors for delivery of genes into neonatal and adult ventricular cardiac myocytes in vitro and in vivo. [J] Basic Res Cardiol 2002;97:348-358
[3]. Mizuguchi H, Koizumi N,et.al. CAR- or alphav integrin-binding ablated adenovirus vectors, but not fiber modified vectors containing RGD peptide, do not change the systemic gene transfer properties in mice. [J] Gene Ther 2002;9:769-776
[4]. Havenga MJ, Lemckert AA,et al. Exploiting the natural diversity in adenovirus tropism for therapy and prevention of disease. [J] J Virol 2002;76:4612-4620
[5]. Wang G, Berk AJ. In Vivo association of adenovirus large E1A protein with the human mediator complex in adenovirusinfected and -Transformed cells. [J] J Virol 2002;76:9186-9193
[6]. Stevens JL, Cantin GT, et al. Transcription control by E1A and MAP kinase pathway via Sur2 mediator subunit. [J] Science 2002;296: 755-758
[7]. Nada S, Trempe JP. Characterization of adeno-associated virus rep protein inhibition of adenovirus E2a gene expression. [J]Virology 2002;293:345-355
[8]. Jing XJ, Kalman-Maltese V,et.al. Inhibition of adenovirus cytotoxicity, replication, and E2a gene expression by adeno-associated virus. [J] Virology 2001;91:140-151
[9]. Pandori M, Hobson D, et.al Adenovirus-microbead conjugatespossess enhanced infectivity: a new strategy for localizedgene delivery. [J]Virology 2002;299: 204-212
[10]. Nasz I, Adam E. Recombinant adenovirus vectors for genetherapy and clinical trials. [J] Acta Microbiol Immunol Hung 2001;48:323-348
[11]. Adam E, Nasz I. Adenovirus vectors and their clinical application in gene therapy. [J]Orv Hetil 2001;142:2061-2070.
[12].赫春秋,冯志华等人,腺病毒载体的特点及其在HCV研究的应用[J].世界胃肠杂志.2003 ;11(6):803-805
[13]. Beltrami AP, Barlucchi L, Torella D,et al .Adult Cardiac Stem Cells Are Multipotent and Support Myocardial Regeneration[J]. Cell, 2003, 114(6):763 -76.
[14]. Coviella CI, Follettie MT, Mellott TJ, et al.Bone morphogenetic protein 9 induces the transcriptome of basal forebrain cholinergic neurons[J]. Proc Natl Acad Sci U S A ,2005 ,102(19):6984-9.
[15]. David L, LamandéN, Gasc JM, et al.Bone morphogenetic protein-9 is a circulating vascular quiescence factor[J].Circ Res. 2008,102(8): 914-22.
[16]. Tang N, Song WX, Luo J, et al.BMP9-induced osteogenic differentiation of mesenchymal progenitors requires functional canonical Wnt/beta-catenin signaling[J].J Cell Mol Med. 2008,[Epub ahead of print]
[1]. Vulliet YS, Greeley M. et al.Intra-coronary arterial injection of mesenchymal stromal cells and microinfarction in dogs. [J] Lancet.2004; 362(9411) :783-784;
[2]. Yoon YS ,Park JS,et al. Unexpected severe calcification after transplantation of bone marrow cells in acute myocardial infaration. [J] Circulation. 2004; 109(25): 3254-3157
[3]. Houghton JM, Stoicov C,et al. Gastric ancer originating from bone marrow derived cells. [J]Science.2004;306(5071):1568-1571
[4]. Yang lei,Mark,H et.al Human cardiovascular progenitor cells develop from a KDR+ embryonic-stem-cell-derived population;[J],Nature 2008 453, 524-528
[5]. Mazhari R Hare JM. Advances in cell-based therapy for structural heart disease;[J] Prog Cardiovasc Dis. 2007 May-Jun;49(6):387-95.
[6].向平,宋银子,陈沅等,提高大鼠心肌梗死模型成功率及存活率的研究。[J]第三军医大学学报,2008;30:4,329-332。
[7]. Ohara F,Sugimoto T, et al. Protective Effect of FR168888, a New Na+/H+ Exchange Inhibitor, on Ischemia and Reperfusion-Induced Arrhythmia and Myocardial infarction in Rats:In Comarison With Other Cardioprotetive Compounds. [J] Jpn.J. Pharmacol, 1999;80:295-302.
[8]. Tousoulis D, Briasoulis A. et.al. Heart regeneration:;what cells to use and how? [J]Curr Opin Pharmacol. 2008 Apr;8(2):211-8. Epub 2008 Mar 4.;
[9]. Zeng Bin, Lin-Guo Sheng. et.al Efficient cardiomyocyte differentiation of embryonic stem cells by bone morphogenetic protein-2 combined with visceralendoderm-like cells.[J] Cell Biology International 2006, 769-776;
[10]. Rebecca Y. Kim, Elizabeth J. Robertson, et al. Bmp6 and Bmp7 Are Required for Cushion Formation and Septation in the Developing Mouse Heart;[J] Developmental Biology ,2001,235, 449–466
[11]. Taha MF, Valojerdi MR,et.al. Effect of bone morphogenetic protein-4 (BMP-4) on cardiomyocyte differentiation from mouse embryonic stem cell; [J] Int J Cardiol. 2007, Aug 9;120(1):92-101.
[12]. Coviella CI, Follettie MT, Mellott TJ, et al.Bone morphogenetic protein 9 induces the transcriptome of basal forebrain cholinergic neurons. [J] Proc Natl Acad Sci U S A ,2005 ,102(19):6984-9.
[13]. David L, LamandéN, Gasc JM, et al.Bone morphogenetic protein-9 is a circulating vascular quiescence factor. [J]Circ Res. 2008,102(8): 914-22.
[14]. Tang N, Song WX, Luo J, et al.BMP9-induced osteogenic differentiation of mesenchymal progenitors requires functional canonical Wnt/beta-catenin signaling. [J] Cell Mol Med. 2008,[Epub ahead of print]
[15]. Buddhadeb Dawn, Adam B. Stein , et al. Cardiac stem cells delivered intravascularly traverse the vessel barrier, regenerate infarcted myocardium, and improve cardiac function;[J] Proc Natl Acad Sci USA, 2005, 102(10):3766-3771
[16].黄枫,余志斌等.心肌肌原纤维的分子结构与功能.[J]心脏杂志;2000,12,(1);
[17].谭金童,陈沅等.BMP9对C3H10T1/2干细胞向心肌样细胞分化的影响[J].重庆医科大学学报,2009,Vol.34 NO.6
[1]. Keiichi Fukuda, Shinsuke Yuasa. Stem Cells as a Source of Regenerative Cardiomyocytes; [J] Circ. Res. 2006 Apr 28;98(8):1002-13.
[2]. Kenneth R. Boheler, Jaroslaw Czyz, David Tweedie, et al. Differentiation of Pluripotent Embryonic Stem Cells Into Cardiomyocytes; [J] Circ. Res. 2002 Aug 9;91(3):189-201
[3]. Kawai T, Takahashi T ,Esaki M, et al. Efficient Cardiomyogenic Differentiation of Embryonic Stem Cell by Fibroblast Growth Factor 2 and Bone Morphogenetic Protein 2;[J]Circ J. 2004 Jul;68(7):691-702
[4]. Li TS, Komota T, Ohshima M,et al .TGF-b induces the differentiation of bone marrow stem cells into immature cardiomyocytes;[J]Biochem Biophy Res Commun.2008 Feb 22;366(4):1074-80
[5]. Susana M. Chuva De Sousa Lopes , Alie Feijen , Jeroen Korving, et al. Connective Tissue Growth Factor Expression and Smad Signaling During Mouse HeartDevelopment and Myocardial Infarction; [J]Dev Dyn .2004 Nov;231(3):542-50
[6]. Laura Lagostenaa, Daniele Avitabileb, Elena De Falcoa ,et al. Electrophysiological properties of mouse bone marrow c-kit+ cells co-cultured onto neonatal cardiac myocytes;[J]Cardiovasc Res. 2005 Jun 1;66(3):482-92。
[7]. Chang Sung-A, Lee EJ ,Kang HJ, et al. Impact of Myocardial Infarct Proteins and Oscillating Pressure on the Differentiation of Mesenchymal Stem Cells: Effect of Acute Myocardial Infarction on Stem Cell Differentiation;[J]Stem Cells .2008 Jul;26(7):1901-12.
[8]. Rajasingh J, Bord E, Hamada H, et al. STAT3-Dependent Mouse Embryonic Stem Cell Differentiation Into Cardiomyocytes Analysis of Molecular Signaling and Therapeutic Efficacy of Cardiomyocyte Precommitted mES Transplantation in a Mouse Model of Myocardial Infarction;[J]Circ. Res. 2007 Oct 26;101(9):910-8.
[9]. Lim JY, Kim WH, Kim J, et al. Involvement of TGF-β1 Signaling in Cardiomyocyte Differentiation from P19CL6 Cells; [J]Mol. Cells, 2007 Dec 31;24(3):431-6
[10]. Choi SC, Yoon J, Shim WJ, et al. 5-azacytidine induces cardiac differentiation of P19 embryonic stem cells; [J]Exp Mol Med, 2004 Dec 31;36(6):515-23.
[11]. Boudewijn P.T. Kruithof, Bram van Wijk, Semir Somi,et al. BMP and FGF regulate the differentiation of multipotential pericardial mesoderm into the myocardial or epicardial lineage; [J] Dev Biol. 2006 Jul 15;295(2):507-22.
[12]. Singla DK, Sobel BE. Enhancement by growth factors of cardiac myocyte differentiation from embryonic stem cells: A promising foundation for cardiac regeneration; [J]Biochem and Biophys Res Commun. 2005 Sep 30;335(3):637-42
[13]. Van Wijk B, Moorman AF, van den Hoff MJ. Role of bone morphogenetic proteins in cardiac differentiation; [J]Cardiovasc Res. 2007 May 1;74(2):244-55.
[14]. Sabine Mazerbourg, Katrin Sangkuhl, Ching-Wei Luo, et al. Identification of Receptors and Signaling Pathways for Orphan Bone Morphogenetic Protein/Growth Differentiation Factor Ligands Based on Genomic Analyses; [J] J Bio Chem. 2005 Sep 16;280(37):32122-32.
[15]. Zhang J, Li L .BMP signaling and stem cell regulation;[J]Devl Bio. 2005 Aug 1;284(1):1-11
[16]. Zeng YA, Rahnama M, Wang S, et al.Drosophila Nemo antagonizes BMP signalingby phosphorylation of Mad and inhibition of its nuclear accumulation ;[J]Development 134, 2061-2071 (2007);
[17]. Shiojioma I, Walsh K. Regulation of cardiac growth and coronary angiogenesis by the Akt/PKB signaling pathway;[J]Genes Dev. 2006 Dec 15;20(24):3347-65 .
[18]. Klaus A, Saga Y, Taketo MM, et al.Distinct roles of Wnt/_-catenin and Bmp signaling during early cardiogenesis;[J]Proc Natl Acad Sci U S A. 2007 Nov 20;104(47):18531-6.
[19]. Papadimou E, Ménard C, Grey C, et al. Interplay between the retinoblastoma protein and LEK1 specifies stem cells toward the cardiac lineage; [J]EMBO J. 2005 May 4;24(9):1750-61.
[20]. Balemans W, Van Hul W. Extracellular regulation of BMP signaling in vertebrates: a cocktail of modulators.; [J]Dev Bio. 2002 Oct 15;250(2):231-50.
[21]. Doss MX, Chen S, Winkler J, et al. Transcriptomic and phenotypic analysis of murine embryonic stem cell der ived BMP2+ lineage cells: an insight into mesodermal patterning; [J] Genome Biol.2007;8(9):R184.
[22]. Taha MF, Valojerdi MR, Mowla SJ. Effect of bone morphogenetic protein-4 (BMP-4) on cardiomyocyte differentiation from mouse embryonic stem cell; [J]Int J Cardiol. 2007 Aug 9;120(1):92-101.
[23]. Dudley AT, Robertson EJ. Overlapping expression domains of bone morphogenetic protein family members potentially account for limited tissue defects in BMP7 deficient embryos; [J]Dev Dyn. 1997 Mar;208(3):349-62.
[24]. Kim RY, Robertson EJ, Solloway MJ. Bmp6 and Bmp7 are required for cushion formation and septation in the developing mouse heart; [J]Dev Biol. 2001 Jul 15;235(2):449-66.
[25]. Somi S, Buffing AA, Moorman AF, et al. Dynamic patterns of expression of BMP isoforms 2,4,5,6, and 7 during chicken heart development. [J]Anat Rec A Discov Mol cell Evol Bio. 2004 Jul;279(1):636-51 .
[26]. Zhang H, Bradley A. Mice deficient for BMP2 are nonviable and have defects in amnion/chorion and cardiac development. [J]Development. 1996 Oct ; 122 (10) :2977-86.
[27]. Winnier G, Blessing M, Labosky PA, et al. Bone morphogenetic protein-4 is required for mesoderm formation and patterning in the mouse. [J]Gene Dev. 1995Sep 1;9(17):2105-16.
[28]. Kingsley DM, Bland AE, Grubber JM, et al. The mouse short ear skeletal morphogenesis locus is associated with defects in a bone morphogenetic member of the TGF beta superfamily. [J]Cell. 1992 Oct 30;71(3):399-410.
[29]. Solloway MJ, Dudley AT, Bikoff EK, Lyons KM, et al. Mice lacking Bmp6 function. [J] Dev Genet. 1998;22(4):321-39.
[30]. Hahn JY, Cho HJ, Kang HJ, et al. Pre-Treatment of Mesenchymal Stem Cells With a Combination of Growth Factors Enhances Gap Junction Formation, Cytoprotective Effect on Cardiomyocytes, and Therapeutic Efficacy for Myocardial Infarction;[J]J Am Coll Cardiol. 2008 Mar 4;51(9):933-43.