摘要
研究采用 1 4～ 1 8d兔胎儿的生殖嵴及周围组织与其同源成纤维细胞共培养 ,低糖DMEM +1 0 %NBS +1 0 %FCS +1 0ng/mLLIF +1 0ng/mLSCF+0 1mol/Lβ 巯基乙醇 +1 0 0U/mL青霉素 +80U/mL链霉素作培养基 ,分离出兔原始生殖细胞 (PGC) ,克隆并多次传代。从原始生殖细胞 (PGC)中获得胚胎生殖细胞 (EG)细胞集落 ,1 4d胎儿原代观察到类EG细胞集落 ,传至 4代后丢失。 1 6d胎儿的类EG只传 2代 ,1 8d胎儿没有得到EG细胞集落。EG细胞具有干细胞的诸多特征 ,呈典型的团块状聚集生长 ,碱性磷酸酶 (AKP)染色呈阳性 ,在衰老饲养层的培养基中生长形成类胚体、上皮细胞。
Primordial germ cells(PGC)were isolated from 14～18 d rabbit embryonic genital ridges and co cultured on homogenous embryonic fibroblast.The medium was low glucose DMEM,supplied with 10%NBS,10%FCS,0 1 mmol/L β mercaptoethunol,100 U/mL penicilin,80 U/mL streptomycin sulfate,10 ng/mL SCF and 10 ng/mL LIF.The rabbit EG like cell lines derived from 14 d PGC were cloned to 4 passages and 16 d PGC were cloned to 2 passages.18 d PGC had no EG clone.EG cells shared many characteristics with mouse ES cell:morphology of stem cell,colonial growth,maintenance of an undifferentiated state through serial passage,ability to form embryonic bodies in suspension culture and multiple type of differentiated cells in monolayer culture.
出处
《畜牧与兽医》
北大核心
2002年第4期1-3,共3页
Animal Husbandry & Veterinary Medicine
基金
省科技厅攻关项目 (0 1 2 2 0 4 1 7)
