不同核移植方法对牛体细胞核移植效率的影响

Effects of Nuclear Transplantation Methods on the Efficiency of Somatic Nuclear Transfer in Bovine

以牛皮肤成纤维细胞为供体细胞 ,比较了电融合法和细胞质内注射法 2种核移植方法对体细胞核移植效率的影响。电融合法构建重组胚的效率显著低于细胞质内注射法 (4 7.1%比 89.0 % ,P<0 .0 1) ;重组胚培养 36 h后的裂卵率和培养 8d时囊胚发育率无明显差异 (76 .4 %比 73.2 % ,11.2 %比 12 .3% ;P>0 .0 5 ) ,但相对于操作的卵母细胞总数而言 ,电融合法得到的总囊胚发育率显著低于细胞质内注射法 (5 .6 %比 10 .9% ,P<0 .0 1)。结果表明 ,用细胞质内注射法进行体细胞核移植效率更高。将发育到桑椹胚和囊胚期的 95枚核移植胚胎移植到 33头受体牛 ,其中 31头受体牛在移植后第 2个情期前返情 ;1头受体牛在移植后 75 d返情 ,但未进行直肠检查 ,无法确定妊娠情况 ;1头受体牛在移植后 6 0 d未见返情 ,直肠检查确证妊娠 ,93d受体牛流产。胚胎移植结果表明 。

Effects of nuclear transplantation methods on the efficiency of somatic nuclear transfer in bovine were investigated.Compared with electric fusion method,intracytoplasmic nuclear injection(ICNI) was more efficient in producing reconstituted embryos when skin fibroblast cells were used as donors(47 1% vs. 89 0%,P<0 01).Although there was no difference between the electric fusion method and ICNI in the cleavage rate(76 4% vs. 73 2%,P>0 05) or blastocyst development(11 2% vs. 12 3%,P>0 05) of reconstituted embryos after cultured for 36 h or 8 days,the percentage of blastocysts relative to total manipulated oocytes was significantly lower in electric fusion method than in ICNI(5 6% vs. 10 9%,P<0 01).These results indicate ICNI is an efficient procedure in bovine somatic nuclear transplantation.Ninety five nuclear transplanted morulae and blastocysts were transferred to 33 synchronous recipients.Out of them,31 recipients showed oestrus before next oestrous cycle and one recipient returned oestrus at 75 days after embryo transfer.One recipient was confirmed pregnant at 60 days by rectal palpation and unfortunately aborted at 93 days after embryo transfer.These results suggested that nuclear transferred embryos derived from adult fibroblast cells in our system have the ability to maintain early pregnancy after transferred to recipients.