人牛异种克隆与牛同种克隆的研究
摘要
本研究分别以人包皮成纤维细胞和牛胎儿成纤维细胞为核供体,牛卵母细胞为受体,构建人-牛异种与牛同种克隆胚。
成功建立了人和牛成纤维细胞系,并对人成纤维细胞F5、F10代细胞做了生长曲线和核型分析,结果表明,基本符合细胞生长规律,且88.5%、82.4%的染色体核型保持正常倍数(2n=64)。
电激活法、化学激活法和精子提取物注射激活法都可激活牛卵母细胞,并使其发育至囊胚。三种方法的最高卵裂率依次为,68.62 %、76.67%、75.44%,囊胚率为11.59%、15.22%、13.95%。
血清饥饿和汇合处理的人成纤维细胞都可支持去核牛卵母细胞的发育,使用不同的电压、融合时间所得最高融合率为58.82%、62.19%,卵裂率为57.50%、60.23%,囊胚率为18.5%、20.1%。
适当延长融合与激活的时间间隔有助于重构胚的发育。
与牛同种克隆胚相比,人牛异种重构胚发育能力低,这种差异是显著的。
In this study ,human foreskin fibroblasts were used as donor cells injected into the enucleated bovine oocytes in nuclear transfer, whereas bovine fetal fibroblasts were used to produce intraspecies embryos.
Human foreskin tissue fibroblasts were dissociated and cultured by explant tissue culture in vitro at 37℃、5% CO2 and 100% humidity; The karytypes show that human fibroblast cell had more normal chromosome content after at least 12 passages(2n=46).
The in vitro matured bovine oocytes were panthenogenetically activated by electroactivation, chemokinesis and sperm-extract injection. The highest cleavage rate were 68.62 %、76.67%、75.44% respectively , and the blastocyst rates were 11.59%、15.22%、13.95%, respectively.
The intraspecies embryos were reconstructed from serum-starved and confluence. The fusion rates were 58.82%、62.19%,cleavage rates were 57.50%、60.23%,and the blastocyst rates were 18.5%、20.1%, respectively.
Compared with bovine intraspecies embryos, human-bovine interspecies embryos had a lower capability of development ,and this difference was significance.
成功建立了人和牛成纤维细胞系,并对人成纤维细胞F5、F10代细胞做了生长曲线和核型分析,结果表明,基本符合细胞生长规律,且88.5%、82.4%的染色体核型保持正常倍数(2n=64)。
电激活法、化学激活法和精子提取物注射激活法都可激活牛卵母细胞,并使其发育至囊胚。三种方法的最高卵裂率依次为,68.62 %、76.67%、75.44%,囊胚率为11.59%、15.22%、13.95%。
血清饥饿和汇合处理的人成纤维细胞都可支持去核牛卵母细胞的发育,使用不同的电压、融合时间所得最高融合率为58.82%、62.19%,卵裂率为57.50%、60.23%,囊胚率为18.5%、20.1%。
适当延长融合与激活的时间间隔有助于重构胚的发育。
与牛同种克隆胚相比,人牛异种重构胚发育能力低,这种差异是显著的。
In this study ,human foreskin fibroblasts were used as donor cells injected into the enucleated bovine oocytes in nuclear transfer, whereas bovine fetal fibroblasts were used to produce intraspecies embryos.
Human foreskin tissue fibroblasts were dissociated and cultured by explant tissue culture in vitro at 37℃、5% CO2 and 100% humidity; The karytypes show that human fibroblast cell had more normal chromosome content after at least 12 passages(2n=46).
The in vitro matured bovine oocytes were panthenogenetically activated by electroactivation, chemokinesis and sperm-extract injection. The highest cleavage rate were 68.62 %、76.67%、75.44% respectively , and the blastocyst rates were 11.59%、15.22%、13.95%, respectively.
The intraspecies embryos were reconstructed from serum-starved and confluence. The fusion rates were 58.82%、62.19%,cleavage rates were 57.50%、60.23%,and the blastocyst rates were 18.5%、20.1%, respectively.
Compared with bovine intraspecies embryos, human-bovine interspecies embryos had a lower capability of development ,and this difference was significance.
引文
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