幼龄杜泊羊胚胎体外生产技术的研究
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摘要
为探讨幼龄杜泊羊胚胎体外生产技术(JIVET),给幼龄杜泊羊卵巢卵泡的利用提供技术支撑,试验采用促卵泡素(FSH)和孕马血清(PMSG)对4~8周龄杜泊羊进行诱导、活体采集卵母细胞、卵母细胞体外成熟-受精-受精卵培养及2~4细胞期受精卵移植研究。结果表明:从8只4~8周龄杜泊羊中采集到可用卵母细胞585枚,平均73.13枚/只;获得2~4细胞期受精卵346枚,卵裂率59.15%;将77枚2~4细胞期体外受精卵移植于17只受体母羊的输卵管壶腹部,7只母羊受孕,产羔13只,受胎率41.18%。说明采用4~8周龄杜泊羊胚胎体外生产技术可作为JIVET的有效技术。
Providing technical support for the utilization of ovarian follicles in young Dorper sheep ovarian follicles in order to study the technology in vitro production of young Dorper sheep embryos. Tests were conducted from 4 to 8-week-old Dorper sheep,live-collected oocytes,oocyte maturation-fertilization-zygote cultivation and 2-4 fertilized ovum transplantation using follicle stimulating hormone(FSH)and pregnant horse serum(PMSG)methods to do Dorper sheep JIVET technology research. The results showed that 585 oocytes were collected from 8 Dorper sheep at the age of 4 and 8 weeks,with an average of 73.13 oocytes. 346 of the2-4 cell fertilized eggs were obtained,whose cleavage rate was 59.15%. 77 in vitro fertilized eggs at the stage of 2-4 cells were transplanted in ampulla of fallopian tube for 17 recipients,in them,7 ewes were pregnant,producing 13 lambs,and the pregnancy rate was 41.18%. It was concluded that the in vitro production of 4 to 8-week-old Dorper sheep embryos could be used as an effective technique for JIVET.
Providing technical support for the utilization of ovarian follicles in young Dorper sheep ovarian follicles in order to study the technology in vitro production of young Dorper sheep embryos. Tests were conducted from 4 to 8-week-old Dorper sheep,live-collected oocytes,oocyte maturation-fertilization-zygote cultivation and 2-4 fertilized ovum transplantation using follicle stimulating hormone(FSH)and pregnant horse serum(PMSG)methods to do Dorper sheep JIVET technology research. The results showed that 585 oocytes were collected from 8 Dorper sheep at the age of 4 and 8 weeks,with an average of 73.13 oocytes. 346 of the2-4 cell fertilized eggs were obtained,whose cleavage rate was 59.15%. 77 in vitro fertilized eggs at the stage of 2-4 cells were transplanted in ampulla of fallopian tube for 17 recipients,in them,7 ewes were pregnant,producing 13 lambs,and the pregnancy rate was 41.18%. It was concluded that the in vitro production of 4 to 8-week-old Dorper sheep embryos could be used as an effective technique for JIVET.
引文
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[2]Rangel-Santos R,Mcdonald M F,Wickham G A.Evaluation of the feasibility of a juvenile MOET scheme in sheep[J].Proceedings of the New Zealand Society of Animal Production,1991,51(1):139-142.
[3]Kelly J M,Kleemann D O,Walker S K.The effect of nutrition during pregnancy on the in vitro production of embryos from resulting lambs[J].Theriogenology,2005,63(7):2020-2031.
[4]Bai J H,Hou J,Guan H,et al.Effect of 2-mercaptoethanol and cysteine supplementation during in vitro maturation on the developmental competence of oocytes from hormone-stimulated lambs[J].Theriogenology,2008,70(5):758-764.
[5]Gou K M,Guan H,Bai J H,et al.Field evaluation of juvenile in vitro embryo transfer(JIVET)in sheep[J].Animal Reproduction Science,2009,112(3/4):316-324.
[6]白佳桦.性成熟前绵羊、牛JIVET技术研究[D].北京:中国农业大学,2008.
[7]汪立芹,何宗霖,林嘉鹏,等.萨福克羔羊卵泡诱导发育效果的研究[J].中国农业大学学报,2015,20(4):141-146.
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[10]马世科,乔海生,马利青,等.幼畜超数排卵及体外胚胎生产技术在青海高原地区的应用研究[J].青海畜牧兽医杂志,2015,45(2):13-14.
[11]陈晓勇,田树军,桑润滋,等.诱导幼羔卵泡发育及体外胚胎生产[J].农业生物技术学报,2008,16(3):456-460.
[12]张兴会,张世伟,宋先忱,等.幼龄母羊超数排卵及卵母细胞成熟效果分析[J].中国畜牧兽医,2011,38(11):136-139.
[13]吴海风,李发弟,常卫华,等.羔羊超数排卵及体外受精技术研究[J].甘肃农业大学学报,2011,46(6):1-5.
[14]李运生,曹鸿国,刘亚,等.羔羊超数排卵与胚胎体外生产技术研究进展[J].动物医学进展,2011,32(11):99-104.
[15]郭宏,万鹏程,代蓉,等.羔羊卵母细胞体外胚胎生产影响因素分析[J].湖北农业科学,2013,52(19):4726-4729.
[16]闵江涛,杜卫华,赵学明,等.绵羊卵母细胞采集、体外成熟和胚胎体外培养对体外受精技术效率的影响[J].中国畜牧兽医,2014,41(5):202-207.
[17]吴伟伟,徐新明,张廷虎,等.影响新疆细毛羊羔羊超排及体外受精效果的研究[J].中国畜牧兽医,2011,38(4):144-147.
[18]于勇,钟旭,王芬露,等.不同月龄绵羔羊卵泡发育的比较研究[J].中国畜牧杂志,2012,48(9):17-21.
[19]郭洪,万鹏程,石文艳,等.EGF和β-巯基乙醇对羔羊卵母细胞体外受精和体外发育的影响[J].安徽农业科学,2012,40(11):6537-6540.
[20]常迪,高雅,罗桂河,等.波尔山羊羔羊超数排卵及体外受精的研究[J].北京农学院学报,2009,24(4):28-31.
[2]Rangel-Santos R,Mcdonald M F,Wickham G A.Evaluation of the feasibility of a juvenile MOET scheme in sheep[J].Proceedings of the New Zealand Society of Animal Production,1991,51(1):139-142.
[3]Kelly J M,Kleemann D O,Walker S K.The effect of nutrition during pregnancy on the in vitro production of embryos from resulting lambs[J].Theriogenology,2005,63(7):2020-2031.
[4]Bai J H,Hou J,Guan H,et al.Effect of 2-mercaptoethanol and cysteine supplementation during in vitro maturation on the developmental competence of oocytes from hormone-stimulated lambs[J].Theriogenology,2008,70(5):758-764.
[5]Gou K M,Guan H,Bai J H,et al.Field evaluation of juvenile in vitro embryo transfer(JIVET)in sheep[J].Animal Reproduction Science,2009,112(3/4):316-324.
[6]白佳桦.性成熟前绵羊、牛JIVET技术研究[D].北京:中国农业大学,2008.
[7]汪立芹,何宗霖,林嘉鹏,等.萨福克羔羊卵泡诱导发育效果的研究[J].中国农业大学学报,2015,20(4):141-146.
[8]郭洪,万鹏程,石文艳,等.不同品种绵羊超数排卵及胚胎体外生产技术研究[J].畜牧与兽医,2012,44(10):20-24.
[9]刘宗正,李和刚,程明,等.利用JIVET技术快速繁育萨福克羊的研究[J].畜牧与饲料科学,2016,37(3):1-4.
[10]马世科,乔海生,马利青,等.幼畜超数排卵及体外胚胎生产技术在青海高原地区的应用研究[J].青海畜牧兽医杂志,2015,45(2):13-14.
[11]陈晓勇,田树军,桑润滋,等.诱导幼羔卵泡发育及体外胚胎生产[J].农业生物技术学报,2008,16(3):456-460.
[12]张兴会,张世伟,宋先忱,等.幼龄母羊超数排卵及卵母细胞成熟效果分析[J].中国畜牧兽医,2011,38(11):136-139.
[13]吴海风,李发弟,常卫华,等.羔羊超数排卵及体外受精技术研究[J].甘肃农业大学学报,2011,46(6):1-5.
[14]李运生,曹鸿国,刘亚,等.羔羊超数排卵与胚胎体外生产技术研究进展[J].动物医学进展,2011,32(11):99-104.
[15]郭宏,万鹏程,代蓉,等.羔羊卵母细胞体外胚胎生产影响因素分析[J].湖北农业科学,2013,52(19):4726-4729.
[16]闵江涛,杜卫华,赵学明,等.绵羊卵母细胞采集、体外成熟和胚胎体外培养对体外受精技术效率的影响[J].中国畜牧兽医,2014,41(5):202-207.
[17]吴伟伟,徐新明,张廷虎,等.影响新疆细毛羊羔羊超排及体外受精效果的研究[J].中国畜牧兽医,2011,38(4):144-147.
[18]于勇,钟旭,王芬露,等.不同月龄绵羔羊卵泡发育的比较研究[J].中国畜牧杂志,2012,48(9):17-21.
[19]郭洪,万鹏程,石文艳,等.EGF和β-巯基乙醇对羔羊卵母细胞体外受精和体外发育的影响[J].安徽农业科学,2012,40(11):6537-6540.
[20]常迪,高雅,罗桂河,等.波尔山羊羔羊超数排卵及体外受精的研究[J].北京农学院学报,2009,24(4):28-31.
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