培养液对猪卵母细胞体外成熟及孤雌胚胎早期体外发育的影响

Effects of Medium on In Vitro Maturation of Pig Oocytes and In Vitro Development of Early Parthenogenetic Embryos

[目的]提高卵母细胞体外成熟质量,优化猪早期胚胎体外培养体系。[方法]以改良TCM199,NCSU-23培养液为基础液,分别添加10%的猪卵泡液(PFF)和10%的优质胎牛血清(FBS)进行卵母细胞体外成熟培养,以成熟率、孤雌胚胎体外发育率等为指标,研究不同培养液对猪卵母细胞体外成熟及孤雌胚胎体外发育的影响。[结果]猪卵母细胞在TCM199,TCM199+FBS和TCM199+PFF组成熟42 h的成熟率分别为(54.2±3.5)%、(68.5±3.2)%和(69.3±3.7)%,在NCSU-23,NCSU-23+FBS和NCSU-23+PFF组成熟42 h的成熟率分别为(51.6±3.3)%、(63.2±3.1)%和(65.5±3.5)%,添加10%的FBS和PFF在0.05水平上显著提高了卵母细胞成熟率;6组不同成熟培养液得到的成熟卵母细胞在孤雌激活后囊胚发育率差异不显著,但TCM199+PFF组的囊胚细胞数(36.5±4.8)在0.05水平上显著高于TCM199和NCSU-23组的囊胚细胞数(18.7±3.2和15.5±2.4)。[结论]改良TCM199,NCSV-23培养液中添加10%PFF和10%FBS可显著促进猪卵母细胞体外成熟及孤雌胚胎早期体外发育。

[Objective] To improve quality of oocytes maturing in vitro and to optimize in vitro culture system of porcine early embryos.[Method] Oocytes were cultured in the improved TCM199 and NCSU-23 basic media,which were added pig follicular fluid（PFF） or high-quality fetal bovine serum（FBS） both at a proportion of 10%（V/V）.After in vitro maturation and development,effects of medium on maturation of pig oocytes and development of early parthenogenetic embryos were investigated using mature rate of pig oocytes and development rate of parthenogenetic embryos as indicators.[Result] After 42 h culture,the mature rates of pig oocytes respectively cultured in the TCM199,TCM199 added FBS,TCM199 added PFF,NCSU-23,NCSU-23 added FBS and NCSU-23 added PFF were（54.2±3.5）%,（68.5±3.2）%,（69.3±3.7）%,（51.6±3.3）%,（63.2±3.1）% and（65.5±3.5）%,respectively.The pig oocytes cultured in the TCM199 or NCSU-23 that was added FBS or PFF had significantly higher mature rate（P0.05）.The development rates of parthenogenetic embryos were not significantly different between these six experimental media.However,the parthenogenetic embryos which developed in the TCM199 added PFF（36.5±4.8） had significantly more blastomeres than those developed in the TCM199 or NCSU-23（18.7±3.2 and 15.5±2.4,respectively）（P0.05）.[Conclusion] The improved TCM199 and NCSU-23 that were added PFF or FBS can largely promote in vitro maturation of pig oocytes and in vitro development of early parthenogenetic embryos.