无血清培养基IVD101和G1/G2在牛体细胞核移植中的应用研究

Effects of in vitro Developmental Competence of Bovine Somatic Cell Nuclear Transfer Embryos in Defined Culture Medium (IVD101 and G1/G2)

通过考察牛体细胞核移植重构胚在无血清培养基(IVD101、G1/G2)条件下培养的囊胚发育率及其质量,从而评估无血清培养基支持牛体细胞核移植重构胚体外发育的能力。采用IVD101和G1/G2对牛体细胞核移植胚胎进行体外培养,并以CR1aa+5%FBS作为对照组。无血清培养基IVD101和G1/G2的囊胚发育率与对照组无显著性差异(41.2%±9.1%、42.2%±10.8%,48.0%±9.2%,P>0.05)。通过囊胚差异染色和冷冻/解冻胚胎存活率分析胚胎质量,发现无血清培养基ICM/Total略低于对照组(31.8%±10.5%、29.5%±11.9%vs.33.0%±14.8%),但无显著性差异(P>0.05);3个组别的囊胚经程序化冷冻/解冻后无血清培养基的存活率(IVD101、G1/G2)略高于对照组,但差异不显著(84.8%、80.4%vs.77.3%,P>0.05)。结果证明无血清培养基(IVD101、G1/G2)可以支持体细胞核移植重构胚的体外发育,且其对程序化冷冻的耐受性与添加血清组(CR1aa+5%FBS)相似。

The purpose of the study was to investigate in vitro development, quality and frozen tolerance of bovine somatic cell nuclear transfer （SCNT） embryos in two defined culture media （IVD101 and G1/G2）. The SCNT embryos were cultured in three media： IVD101, G1/G2, and CRlaa＋5%FBS （control group）. The results indicated that the development rates of SCNT embryos cultured in IVD101 and G1/G2 were lower than that in CRlaa＋5%FBS, but had no significant difference among three groups （41.2%±9.1%, 42.2%±10.8% and 48. 0%±9.2%, respectively,P〉0.05）. The result of differential staining showed that the ratio of ICM/Total of blastocysts cultured in IVD101 and G1/G2 were lower than that in the group of CRlaa＋5% FBS, whereas there was no significant difference among them （31.8 ±10.5 %, 29.5 % ± 11.9 % and 33.0%±14.8%, respectively, P〉0.05）. No significant difference was observed in the survival rate among three groups （84.8%, 80.4% and 77.3%, respectively, P〉0.05）. Our study demonstrated that defined cultured media （IVD101 and G1/G2） could support in vitro develop- ment of bovine SCNT embryos, and freezing tolerance of them were similar with the culture medium with FBS.