牛胚胎生殖细胞的分离与培养

Isolation and culture of bovine embryonic germ cells

胚胎生殖细胞 (Embryonicgermcells,EG)是由生殖嵴原始生殖细胞 (Primordialgermcells,PGCs)中分离得到的一种未分化而多潜能的干细胞。牛EG细胞的研究在EG细胞核移植、转基因及建立生物反应器方面具有广阔的应用前景。本研究从 2 9- 70日龄牛胎儿PGCs分离得到EG细胞 ,经过抑制分化培养 ,其中一个细胞系传至 6代。所分离得到的EG细胞具有典型的EG细胞形态 ,AP及PAS染色呈阳性 ,核型正常 ,同时观察到这些细胞在体外进行自发性分化 ,可形成类胚体。

Embryonic germ (EG) cells are undifferentiated pluripotential stem cells derived from primordial germ cells (PGCs) of the genital ridge. The study on bovine EG cells has great value and promising prospects in nuclear transfer, transfer gene and establishment of bio-reactor of EG cells. In this study, bovine EG cells were isolated from PGCs of 29-70 d fetuses aborted and cultured undifferentiatedly on mitomicin-inactivated mouse STO or MEF feeders. One bovine EG-like cell line had been successively maintained for 6 passages. The pluripotent nature of EG cells was identified by typical cellular and colonial morphology of EG cells, positive AKP staining and positive PAS staining, and normal karyotype. These EG-like cells can also spontaneously differentiate into embryoid bodies, fibroblasts-like cells and neuron-like cells in vitro according to observation .