从原始生殖细胞分离和克隆兔的胚胎生殖细胞  

Isolation and clone of embryonic germ cell from rabbit primordial germ cell

作　　者：桑润滋[1] 周振明[1] 韩建永[1] 孙国杰[1] 李俊杰[1] 

机构地区：[1]河北农业大学动物科技学院,河北保定071001

出　　处：《畜牧与兽医》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)

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.

关 键 词：分离 克隆 原始生殖细胞 胚胎生殖细胞 兔 胚胎发育 胚胎干细胞 

分 类 号：Q132.4[生物学—普通生物学]