山羊原始生殖细胞的分离与培养

从46例山羊胎儿中分离培养原始生殖细胞,发现胎龄在25～38d的胎儿原代培养时都可获得大量的细胞集落,适合做胚胎生殖细胞的分离培养,最高传至6代。以小鼠胎儿成纤维细胞(MEF)、山羊胎儿成纤维细胞(GEF)、牛胎儿成纤维细胞(BEF)为饲养层,在不添加细胞因子情况下,都可以培养出原代山羊的胚胎生殖细胞。传代结果表明,MEF的培养效果较好,但与GEF组差异不显著(P>0.05),BEF组培养效果较差(P<0.05)。以3个不同质量浓度LIF的培养液培养山羊原始生殖细胞(PGC)结果表明,在原代培养时,效果差异不显著(P>0.05);而在传代过程中,以含LIF10ng/mL组培养效果最好,但与含LIF5ng/mL组差异不显著(P>0.05),LIF1ng/mL组培养效果较差(P<0.05)。

Isolation and culture of goat embryonic germ cells

The primordial germ cells were isolated from 46 goat fetuses with embryonic age from 25 to 38 days.The research showed there were many colonies in primary culture.One EG cell line maintained undifferentiation for 6 passages.When goat primordial germ cells were cultured on different feeder cells,MEF,GEF and BEF all could sustain goat primary embryonic germ cells,and MEF was finer than others.But there were no significant difference between MEF and GEF(P>0.05).Goat primordial germ cells were cultured with the medium of 3 different concentrations of LIF.There was no difference in primary culture (P>0.05).When passed,the group containing ng/mL LIF was finer than others.But there was no difference with the group containing 5 ng/mL LIF (P>0.05).The group containing 1 ng/mL LIF was the worst.