自原始生殖细胞分离克隆小鼠ES细胞研究初报

A preliminary study on the isolation and cloning mouse embryonic stem cells derived from primordial germ cells

自交配后 9.5～ 13.5 d小鼠胎儿生殖嵴 /腺或其类似物及周围组织 ,采用与其同源胎儿成纤维细胞共培养的方式分离克隆到具小鼠 ES细胞诸多特征的胚胎生殖细胞 ( EG细胞 )系。即具有连续传代的能力 ( 1例来自交配后 11.5 d胎儿类 ES细胞传至 10代 ) ,部分细胞集落呈典型鸟巢状结构 ,AP染色呈阳性 ,体外分化或延迟传代、堆叠培养具有分化形成类胚体、上皮细胞、成纤维细胞或神经细胞等的能力。同样培养交配后 7.5～ 8.5 d和14.5～ 15 .5 d的小鼠胎儿 ,原代观察到类 ES细胞集落 ,继代培养未观察到类 ES细胞集落 ,16 .5～ 18.5 d小鼠胎儿原代培养未观察到

Embryonic germ cell lines were isolated and cloned from gonad,gonadal ridge,analogue or the surrounding tissues of 9.5～13.5 days postcoitum(dpc) mouse fetuses.Primordial germ cells were co cultured with mouse embryonic fibroblasts obtained from the same conceptus.These cells exhibit features similar to those of murine ES cells such as typical morphology (nest like).These cells have the ability to be continuously passaged.They continue to be AP positive and have the capacity of forming embryoid bodies or differentiating into epithelia like,fibroblast like or neural like cells when in vitro differentiation,delaying passage or growing beyond confluence on fibroblast feeder layers.ES cell like colonies are observed at primary passage of 7.5～8.5 d and 14.5～15.5 d mouse fetus and disappeared at later passage.No ES cell like colonies are observed at primary passage of 16.5～18.5 d mouse fetus.