摘要
目的 :分别使用布法罗大鼠肝细胞 (buffaloratsliver,BRL)条件培养基或鼠胚成纤维细胞 (mouseembry onicfibroblast,MEF)饲养层体外培养 ,比较不同培养条件对胚胎干细胞 (embryonicstemcells,ESC)分化为心肌细胞的影响。方法 :用BRL条件培养基或MEF饲养层抑制ESC分化并保持其增殖 ,以维甲酸 (retinoicacid ,RA)、二甲基亚砜 (DMSO)、转化生长因子 β1(TGF β1)、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 :BRL及MEF饲养层两种条件均能促进ESC增殖 ,6种分化条件均能使ESC分化为心肌细胞 ;其中以BRL条件培养基培养 ,TGF β1(2ng·ml-1)、activin A(2 0ng·ml-1)及 2 0 %胎牛血清 (fetuscalfserium ,FCS)组成的分化培养基可以使 (92 .8± 2 .2 ) %的ESC分化为心肌细胞 ,分化比率显著高于其它各组 (P <0 .0 1)。结论 :以BRL条件培养基培养 ,TGF β1、activin A细胞因子组合是一种理想的ESC体外分化体系。
Objective: To evaluate the different conditions inducing mouse embryonic stem cells (ESCs) in vitro to differentiate into cardiomyocytes. Methods: BRL (Buffalo rats liver ) conditioned medium or MEF (mouse embryonic fibroblast) feeders were used to promote the growth of ESCs and maintain them in an undifferentiated state. During the inducing process,RA,DMSO, TGF-β_1 and activin-A were used as inducing reagents to make up six kinds of differentiating medium. Then a three-step method inducing ESCs cultured in hanging drops,in suspension and in plating was used to induce the differentiation of ESCs. Results: ESCs could be induced in vitro to differentiate into cardiomyocytes. Of all groups,the highest differentiating ratio was obtained in the group induced by TGF-β_1 and activin-A. Conclusion: The system made up of BRL conditioned medium and the inducing reagents including TGF-β_1 and activin-A is valuable in inducing ESCs to differentiate into cardiomyocytes.
出处
《武汉大学学报（医学版）》
CAS
2004年第4期383-386,T003,共5页
Medical Journal of Wuhan University
关键词
胚胎干细胞
心肌细胞
分化
布法罗大鼠肝细胞
成纤维细胞
embryonic stem cells
cardiomyocytes
differentiation
Buffalo rats liver
embryonic fibroblast
