比较体外不同条件诱导胚胎干细胞分化为心肌细胞的实验研究

Comparison of different conditions inducing embryonic stem cells in vitro to differentiate into cardiomyocytes

目的比较体外不同分化条件下胚胎干细胞(ESC)分化为心肌细胞的分化比率。方法用布法罗大鼠肝细胞(BRL)条件培养基抑制ESC分化及保持其增殖,以维甲酸(RA)、二甲基亚砜(DMSO)、转化生长因子β1(TGF-β1)及激活素-A(activin-A)为分化诱导剂,组合成6种分化培养基,采用悬滴悬浮、贴壁三步法诱导ESC分化为心肌细胞,比较各组分化比率。结果6种分化条件均能使ESC分化为心肌细胞,其中以TGF-β1(2μg/L)、activin-A(20μg/L)及20%胎牛血清(FCS)组成的分化培养基可以使高达9280%±222%的ESC分化为心肌细胞,分化比率显著高于其它各组(P<001)。结论以BRL条件培养基培养,TGF-β1、activin-A作分化诱导剂可作为一种理想的ESC体外分化体系。

AIM: To evaluate the different conditions inducing mouse embryonic stem cells (ESC) in vitro to differentiate into cardiomyocytes. METHODS: BRL conditioned medium was used to promote the growth of ESC and maintain them in an undifferentiated state. During the inducing process, retinoic acid (RA), DMSO, activin-A and TGF-β 1 were used as inducing reagents, and made up six kinds of differentiating medium. Then a three-step method inducing ESC cultured in hanging drops, in suspension and in plating was used to induce the differentiation of ESC. RESULTS: ESC were induced in vitro to differentiate into cardiomyocytes. Of all groups, the highest differentiating rate was observed in the group induced by activin-A (20 μg/L) and TGF-β 1 (2 μg/L). CONCLUSION: The inducing conditions including activin-A (20 μg/L) and TGF-β 1 (2 μg/L) is very valuable in inducing ESC differentiation into cardiomyocytes. [