用KOSR优化山羊克隆胚的培养体系

Optimization of Culture Technique for Goat Embryos Cloning with KOSR

体细胞核移植重构胚的体外培养一直是影响山羊克隆效率的一个重要因素,为了排除成分复杂的胎牛血清(fetal bovine serum,FBS)对克隆重构胚发育的潜在抑制作用,本研究采用成分明确的血清替代品(knockout serum replacement,KOSR)进行山羊体细胞克隆重构胚胎的体外培养,并比较KOSR与FBS对重构胚的发育效率的影响。结果显示,KOSR组山羊重构胚培养体系的分裂率明显高于FBS组(80.67±0.63%.vs 49.02±4.85%,P<0.01),2组在重构胚重编程过程中的8细胞率具有显著差异(48.89±1.92%vs.28.59±3.13%,P<0.05);2组不同培养体系的受孕率也有明显不同(33.3%vs.10%,P<0.05)。研究结果表明,KOSR培养体系能有效提高山羊重构胚的发育效率,特别是有利于重构胚的早期重编程,为获得健康的转基因克隆山羊奠定了良好的基础。

The in vitro culture of cloned embryos established by nuclear transplantation is a key factor affecting the efficiency of goat cloning system. To develop an effective culture system for cloned goat embryos,knockout serum replacement （KOSR） was used to take over fetal bovine serum （FBS） in an in vitro culture system. The developmental potential and stability of the reconstructed embryos were then compared. The results showed that the cleavage rate from KOSR was significantly higher than that from FBS （80. 67 ± 0. 63% vs. 49.02 ± 4. 85%,P〈0. 01）. The 8-cell formation rate in KOSR cultured group was much higher than that in FBS （48. 89± 1. 92% vs. 28. 59±3. 13%, P〈0. 05）. In addition, the pregnancy rate was significantly different between the two media groups （33. 3% vs. 10 %, P〈0.05）. This study demonstrated that using KOSR can benefit early reprogramming of cloned embryos and increase the development rate of goat cloned embryos. The culture system in this study greatly increased the efficiency for goat cloning and provides a valuable foundation for generating healthy cloned animals in the future.