兔核移植胚胎的克隆研究

THE STUDY ON RABBIT EMBRYO CLONING BY NUCLEAR TRANSPLANTATION

本研究改进了兔受体卵母细胞的去核程序及供体卵裂球的处理方法，并对核移植胚的体外克隆、继代克隆及克隆胚的体内发育进行了试验。结果表明：卵龄对受体卵母细胞的去核具有显著的影响，卵龄为１５～１８ｈ的去核率为１００％（４５／４５），显著高于１３～１４ｈ的４６．６％（７／１５），Ｐ＜０．０１。ＤＮＡ合成抑制剂Ａｐｈｉｄｉｃｏｌｉｎ处理１６－细胞期卵裂球后，重组胚的囊胚发育率５４％（２０／３７）高于未处理组的４５％（１４／３１），Ｐ＜０．０５。从２枚１６－细胞供体胚分别获得来自一个供体胚的９枚和７枚核移植克隆囊胚，囊胚发育率为５１．６（１６／３１）。然而第一次继代核移植胚的囊胚发育率仅为１１．５％（６／５２）。１６－及３２－细胞期卵裂球的重组胚可发育至产仔，共获２窝８只仔兔。其中１只、２只、２只和３只仔兔分别来自４个供体胚。

In tbis study,the procedures for oocyte enucleation and donor blastomere treat- ment were modified. In vitro recycling of cloned nuclear transplant embryos and theirdevelopment were tested. 15～18 hours post HCG treatment oocytes were enucleated at ahigher rate(100%)than that of 13～14 hours post HCG treatment oocytes(46.6%,P<0.01).The blastocyst development rate(54%)of nuclear transplant embryos fromthe blastomeres treated with Aphidicolin(DNA synthesis inhibitor)medium was higherthan that of the non-Aphidicolin treated group(45%,P<0.05).9 and 7 cloned blas-tocysts were obtained respectively from two donor embryos of 16 cell stage,the per-centage of developing to blastocyst was 51.6(16/31).However,only 11.5%(6/52)ofthe first generation transplant embryos developed to blastocysts.Transfer of nucleartransplant embryos from 16 or 32 cell donor embryos to two recipient rabbits led to thedevelopment of 8 offspring.One,two,two and three of them were respectively from 4donor embryos.