诱导体外培养小鼠胚胎干细胞分化为心肌细胞方法的研究

A method of inducing mouse embryonic stem cells cultured in vitro to differentiate into cardiomyocytes

目的 :采用布法罗大鼠肝细胞条件培养基代替重组LIF ,建立在不加任何化学诱导剂的情况下诱导体外培养小鼠胚胎干细胞 (ES细胞 )分化为心肌细胞的方法。方法 :在诱导分化实验前 ,应用BRL条件培养基来维持ES细胞生长和抑制其分化 ,并采取将悬滴培养法和聚集培养法结合起来 ,分三步诱导ES细胞分化的方法。结果 :分化细胞中可见节律性收缩的细胞 ,经超微结构分析和免疫细胞学鉴定为心肌细胞。结论 :本法无需添加化学诱导剂 ,为一种简便、经济的诱导体外培养小鼠ES细胞分化为心肌细胞的方法。

AIM: To evaluate the method of inducing mouse embryonic stem (ES) cells in vitro to differentiate into cardiomyocytes without using any chemical reagents.METHODS: BRL conditioned medium was used to promote the growth of ES cells and maintain them in an undifferentiated state before the experiment of differentiation. Then a three-step method including ES cell culture in hanging drops and in suspending was used to induce the differentiation of ES cells. RESULTS: Rhythmically contracting cells were observed among differentiated cells, which were proved to be cardiomyocytes with electron microscope and immunocytochemistry. CONCLUSION: A simple and economical method was established to induce mouse ES cells cultured in vitro to differentiate into cardiomyocytes without using any chemical reagents. [