绵羊卵泡液和次黄嘌呤对卵母细胞体外减数分裂的影响

The Effects of Follicular Fluid and Hypoxanthine on Resumption of Meiosis in Ovine Oocytes Cultured in vitro

目的 利用在培养液中添加绵羊卵泡液和次黄嘌呤 ,抑制卵母细胞GVBD发生 ,延长转录活性 ,从而使卵母细胞真正成熟 ,提高胚胎质量及生产效率。方法 利用体外成熟技术对有屠宰采集的绵羊卵母细胞进行培养 ,培养液中添加卵泡液及次黄嘌呤 ,检查成熟效果。结果 将卵母细胞培养在 5 0 %和 10 0 %的卵泡液中 ,2 4h后处于GV期的卵母细胞分别为 19% (8 4 2 )和 33 3% (13 39)。在含有 4mmol L次黄嘌呤的培养液中 ,2 4h后有2 1 6 % (16 74 )的卵母细胞处GV期 ,而对照组中只有 6 % (3 5 0 ) ,经过次黄嘌呤处理的卵母细胞多数都停滞于PⅠ期(44 6 % ,33 74 )。在 4mmol L次黄嘌呤培养液中添加FSH并未使受到抑制的卵母细胞诱导成熟。结论 卵泡液和次黄嘌呤只能在有限的程度上抑制减数分裂的重新启动 ,并对减数分裂的全过程都有影响 ,这种影响程度与抑制因子的浓度相关 ,存在明显的剂量效应。

ObjectiveThe follicular fluid (oFF) and hypoxanthine were added in maturation medium to prevent the GVBD occur,prolong the transcription of the oocyte genome, improve the maturation level of oocyte cytoplasm, enhance the IVF embryo developmental potential. Method The oocytes recovered from the ovaries collected in abattoir were cultured in vitro maturation medium containing follicular fluid(oFF) and hypoxanthine in different concentration. The nuclear phases were checked after 24hrs. Results Oocytes were incubated in culture medium with 50% and 100% follicular fluid for 24h, and 19% and 33 .3% of them were maintained in GV stage, respectively. At a concentration of 4mM hypoxanthine in culture medium, the meiosis resumption of ovine oocytes was inhibited significantly. 21 .6% of oocytes were maintained in GV stage, and 44 .6% were in PI stage. But the inhibitory effect could not be reversed by the induction of FSH. Conclusion The concentration of inhibitory factors had liner relationship with meiosis arrest. There were dose -dependent effects in the control of meiosis.