从原始生殖细胞分离昆明小鼠EG细胞及其传代培养

为提高昆明小鼠胚胎生殖细胞(embryonic germ cells, EG cells)建系效率，以怀孕8.5～12.5 d小鼠胎儿为材料，比较了胎儿后1/3部位的组织共培养、生殖嵴共培养、差速贴壁和穿刺生殖嵴4种分离胚胎原始生殖细胞(primordial germ cells，PGCs)的方法以及不同传代方式对EG细胞克隆的影响。结果表明：生殖嵴共培养和差速贴壁方式获得了较好分离EG细胞的效果，与其它两组比较差异显著；手工传代方式与连同成纤维细胞一起消化的传代方式相比获得了较高传代比率。对所分离EG细胞经形态观察、AKP染色和体外分化能力检测，证实其符合小鼠EG细胞的集落状生长、细胞未分化特性及细胞多能性等特征。

Isolation of Kunming Mice Embryonic Germ Cells Derived From Primordial Germ Cells and Their Passage Culture

Four isolation methods: co-culture back 1/3 position of embryonic tissue, co-culture genital ridges, different speed attaching and puncture the genital ridges were compared to improve the efficiency for establishing the embryonic germ cells (EG cells) line from Kunming mice. Two passage ways, hand and digestion together with fibroblast, were used to clone EG cells. Results showed that co-culture the genital ridges and different speed attaching were more effective methods than that of the others. And passage by hand could improved passage efficiency of EG cells compared with digestion together with fibroblast. When isolated EG cells were examined by morphology observation, AKP strain and in vitro differentiation ability, they were convinced to have a series of characters of mice EG cells including cloning grow way, undifferentiation state and pluripotency of cells.