胰岛素和白血病抑制因子对猪卵母细胞体外成熟和孤雌激活胚胎的影响

Effect of insulin and leukemia inhibit factor on in vitro maturation and parthenogenetic development of porcine oocytes

为探讨胰岛素(Insulin)和白血病抑制因子(Leukemia inhibit factor,LIF)对猪卵母细胞体外成熟(IVM)和猪孤雌激活胚胎(PAEs)的影响,在卵母细胞体外成熟或者胚胎培养基中添加Insulin和LIF,研究卵裂率和囊胚率的变化。结果:添加了5μg/mL Insulin后猪卵母细胞体外成熟效果显著提高,但成熟后孤雌激活发育能力与非添加组相近;而胚胎培养基中添加Insulin对孤雌胚的卵裂和囊胚的形成也没有明显促进作用;添加1 000 U/mL的LIF后,卵母细胞核成熟率没有明显提高,反而孤雌激活后囊胚率急剧下降,但对卵裂率以及囊胚总细胞数影响不大;在胚胎培养基中添加LIF后,孤雌胚的卵裂和囊胚形成并没有明显的提高。表明:Insulin对卵母细胞体外成熟有益,但是对孤雌胚胎的最佳处理程序还需要摸索;本文所采用的LIF处理对猪卵体外成熟以及孤雌胚胎体外发育没有帮助,还需要进一步研究其他浓度和处理程序对猪卵母细胞体外成熟和孤雌激活胚胎发育能力的影响。

To evaluate the effects of supplementation of Insulin and leukemia inhibit factor （LIF） in culture media on in vitro nuclear maturation of porcine oocytes and in vitro development of porcine parthenotes. 1 ） Insulin or LIF was added into the medium of porcine oocytes in vitro maturation; and the matured oocytes with the first poly body were recorded and parthenogentieally activated followed by being cultured in vitro in normal medium; 2） Insulin or LIF was supplemented into parthenotes in vitro culture medium; and the rates of cleavage and hlastocyst formation were recorded. For in vitro maturation, supplementation of 5μg/mL insulin significantly increased the rate of in vitro nuclear maturation of pig oocytes although subsequent in vitro development of these oocytes was similar to the control after parthenogenetic activation. When LIF was added, no obvious increase of nuclear maturation was found whereas the blastocyst rate decreased drastically of such oocytes after parthenogenetic activation. When insulin was only added to culture medium of parthenotes, no beneficial effect was observed regarding to cleavage and blastocyst formation. When LIF was singly supplemented to embryo cuhure medium, very slight, and not significant increase of the cleavage and blastocyst rates was found. Insulin was confirmed conducive to the maturation of porcine oocytes, but further studies are required to investigate if other concentrations or treatments of insulin are helpful to porcine parthnotes development in vitro. Regarding to LIF, it is harmful to blastoeyst development in vitro. However, more concentrations and treatments of LIF are deserved to be examined whether it could promote maturation and in vitro development of porcine embryos or not.