不同时期鸡胚原始生殖细胞分离的研究

ISOLATION AND CULTURE OF PRIMORDIAL GERM CELLS FROM CHICKEN EMBRYOS

采用Ficoll密度梯度离心,酶解离两种方法在鸡胚孵化的第14期、19期、28期,分离、培养鸡胚中的原始生殖细胞(PGCs)。探索PGCs分离、培养的适宜时期及方法,以期获得较多数量,较高活力的PGCs作介导生产转基因鸡。结果表明:1.提取、分离PGCs的最佳时期依次为19期、28期。2.两种分离方法均能分离到一定数量的PGCs细胞。但在19期和28期,酶解离法分离到的PGCs的相对数量较多,存活时间较长,是一种较适宜的分离方法。

Along with two methods of Ficoll density-gradient centrifugation and EDTA-Trypsin collection of isolating primordial germ cells (PGCs) from chicken embryos at three different stages: from blood at stage 14, from genital ridge at stage 19 and gonadal at stage 28. PGCs were then cultured in the TCM-199 medium containing 10% FCS in vitro. The number of viable cells were determined by using the Trypan Blue test; PGCs were identified using PAS reaction. The result showed that, EDTA-Trypsin collection was more preferable to PGCs collection at stage 19 and stage 28, and these two stages were suitable for collection of PGCs from early chicken embryos.