不同类型供体细胞对牛体细胞重构胚发育能力的影响

通过显微操作去除体外成熟培养22￣23h卵母细胞的细胞核,并通过细胞质内注射法将供体细胞核移入去核卵母细胞内构建重组胚,重组胚经离子霉素(Ionomycin)激活5min后,再在6-DMAP内培养4h,将重组胚转CR1培养液中进行培养,24h和36h检查卵裂率,培养8d后统计囊胚发育率。比较了经血清饥饿和未饥饿牛成纤维细胞、颗粒细胞作为供体细胞构建的重组胚。研究发现:未饥饿组的牛成纤维细胞重组胚囊胚发育率(11.76%)显著高于血清饥饿组重组胚囊胚发育率(3.16%,p<0.05)。未饥饿组颗粒细胞囊胚发育率(14.94%)也显著高于血清饥饿组(7.14%,p<0.05)。经血清饥饿处理的颗粒细胞和成纤维细胞重组胚发育率差异不显著(p>0.05),未饥饿处理组两种细胞的重组胚发育率差异不显著。结果表明,用非饥饿处理牛成纤维细胞及颗粒细胞重组胚的发育率较高,牛颗粒细胞更有利于重新程序化。

Effect of Type of The Donor Cell on The Development of Bovine Somatic Cell Nuclear Transplantation Reconstituted Embroys

We studied the type of donor cell on the development of bovine somatic cell nuclear transplantation reconstituted embryos. The oocyte was cultured for 22~23h to reach maturity in vitro. The nuclear of the mature oocyte was taken out by micromanipulation. The donor nuclei came from fibroblasts and granular cells. Cells were divided into two halves, which were respectively treated, by serum starvation and non- starvation. The donor nuclei were transplanted into the enucleated oocyte by the microinjection of cytoplasm to form reconstituted embryos. After being activated in Ionomycin media for 5 min, the recombinant embryos were firstly put into 6-DMAP media to be cultured for 4 h, then into the CR1 media. The cleavage rate of the reconstituted embryos was examined after 24h and 36h and the growth rate of the blastocyst was counted after 8 d. The results show that the development rate of blastocyst of the somatic cell treated by serum starvation is strikingly higher than that of the reconstituted embryos from bovine somatic cell treated by serum starvation.(fibroblasts: 3.16% vs 11.76%, p<0.05; granular cells: 7.14% vs 14.96%, p<0.05).There is not much difference in the development rate of blastocyst between the reconstituted embryos from fibroblasts and granular cells. The result implies that the reconstituted embryos from non- starvation treated somatic cells have higher development rate and the embryos from bovine granular cells have higher development rate in vitro.