大鼠心肌条件培养基在昆明白小鼠胚胎干细胞分离培养中的效果

Effect on isolation and culture of Kunming white mouse ES cells with media conditioned of rat cardiomyocyte

采用大鼠心肌条件培养基对昆明白小鼠胚胎干细胞进行分离、消化传代、培养鉴定等,研究大鼠心肌条件培养基在昆明白小鼠胚胎干细胞分离培养中的作用效果。结果显示,SD大鼠心肌条件培养基在昆明白小鼠3.5d囊胚胚胎干细胞集落分离中,所分离的昆明白小鼠胚胎干细胞集落具有岛屿状生长,隆起于饲养层上;碱性磷酸酶染色为强阳性;体外分化可形成类胚体状结构,贴壁的类胚体周边会出现许多分化的上皮样细胞或单个散在的细胞;经过常规冻存传代的胚胎干细胞集落具有较为一致的生长形态,并呈现胚胎干细胞集落所特有的岛屿状生长形态;对传至第5代的小鼠胚胎干细胞集落进行核型分析,小鼠胚胎干细胞核型正常率大于75%。试验证实,SD大鼠心肌条件培养基在昆明白小鼠胚胎干细胞分离培养过程中效果较好。

Effect on isolation and culture of Kunming white mouse embryonic stem cells with media conditioned of rat cardiomyocyte was studied in this paper by isolation,digestion,passage,culture,identification and other ways.Results show that mouse embryonic stem cell clusters isolated were island-like on feeder layers of mouse embryonic fibroblast;Alkaline phosphatase staining of mouse embryonic stem cell was strongly positive;Mouse embryonic stem cells could become embyonic bodies-like structures in vitro,and embryonic bodies which stuck to plates could be differentiated into epithelial-like cells and scattered single cell;Embryonic stem cells were identical and island-like image by route freeze and culture;Karyotype analysis to the 5th passage embryonic stem cells was researched,and the normal rate was over 75%.It indicated that the effect of media conditioned of SD rat cardiomyocyte to isolation and culture of Kunming white mouse embryonic stem cell was very good.