CB·氨基酸在猪卵母细胞孤雌激活及体外培养中的作用

Effect of CB and Amino Acids on Parthenogenetic Activation and in vitro Culture of Porcine Oocytes

[目的]研究CB、氨基酸在猪卵母细胞孤雌激活及体外培养体系中的作用,为优化相关技术体系提供依据。[方法]卵母细胞经体外成熟培养,采用不同孤雌激活方法(Ele.+CB组、CB组、Ele.组、对照组),研究CB在激活中的作用。激活后采用PZM3培养液,并去除氨基酸成分,探索氨基酸对体外培养胚胎的作用。[结果]CB+Ele.组的卵裂率显著地高于其他3组(P<0.05),囊胚率为32.7%,囊胚细胞数为61.4,而其他3组均未出现囊胚。电激活(脉冲电压100 V/mm,脉冲时程100μs,脉冲次数1次)联合CB处理,可激活卵母细胞体外发育至囊胚,CB有助于提高卵裂率、囊胚率。添加氨基酸的培养液中的卵母细胞的卵裂率、囊胚率、囊胚细胞数显著提高(P<0.05),表明氨基酸能提高猪孤雌激活胚胎培养效果。[结论]添加CB、氨基酸在猪卵母细胞孤雌激活及体外培养体系中能提高胚胎的培养效果。

[Objective] To investigate the effect of CB and amino acids on parthenogenetic activation and in vitro culture of porcine oocytes,and offer the basis to optimize the early embryo culture system of porcine.[Method] After maturation of porcine oocytes in vitro,different parthenogenetic activation methods（Ele.＋CB group,CB group,Ele.group and control group） were used to explore the effect of CB.Parthenogenetic oocytes were put into the PZM3 culture medium without amino acids to explore the effect of amino acids.[Result] The cleavage rate of Ele.＋CB group was significant higher than that of other groups（P〈0.05）.The blastocyst rate of Ele.＋CB group was 32.7% and the number cells of blastocyst were 61.4,but other three groups didn＇t develop to blastocyst stage.The condition of electro-activation（100 V/mm pulse voltage,100 μs pulse duration and 1 pulse time） combined with CB could activate oocytes to develop to blastocyst and CB contributed to the improvement of cleavage rate and blastocyst rate.The cleavage rate,the blastocyst rate and the number of blastocyst cells in culture medium with amino acids were significantly higher than that in medium without amino acids（P〈0.05）.The result showed that amino acids could improve the development of porcine parthenogenetic embryos in vitro.[Conclusion] In conclusion,CB and amino acids had positive effect on parthenogenetic activation and in vitro culture of porcine oocytes.