抗坏血酸体外诱导人胚胎生殖细胞向心肌细胞分化  

Ascorbic acid induces differentiation of human embryonic germ cells towards cardiomyocytes

作　　者：陈永珍[1] 朱旻[1] 李芳[1] 张于娟[1] 

机构地区：[1]苏州大学医学部解剖学与组织胚胎学系,215123

出　　处：《江苏医药》2010年第13期1551-1554,F0002,共5页Jiangsu Medical Journal

基　　金：苏州大学医学发展基金项目(EE134710)

摘　　要：目的探讨抗坏血酸作为诱导剂诱导人胚胎生殖细胞(hEGC)体外分化为心肌细胞。方法取5～10周人胚胎生殖腺嵴,进行组织块体外培养,用直接悬浮法使hEGC形成拟胚体(EBs),用不同浓度抗坏血酸的培养基对其进行诱导分化。然后取不同时间的细胞做免疫细胞化学染色,鉴定细胞的心肌特异转录因子GATA-4和心肌肌钙蛋白T(cTnT)表达情况。结果抗坏血酸诱导hEGC分化为心肌细胞的最佳浓度为0.1mg/ml,诱导第3周时分化率达(85.2±1.8)%,显著高于不添加任何诱导剂的对照组。诱导后细胞形态变成梭形,4周时细胞突起相互连接成条索状,且排列方向趋于一致;诱导后1周即开始出现GATA-4弱表达,第3周时表达最强;诱导后2周,细胞内开始表达cTnT,3周强阳性表达,4周表达明显增强。结论抗坏血酸能够促进hEGC向心肌细胞分化,可用以建立体外诱导hEGC分化为心肌细胞的方法 。Objective To investigate the feasibility of differentiation of human embryonic germ cells（hEGCs） towards cardiomyocytes induced by.ascorbic acid.Methods hEGCs of 5 to 10 weeks old were taken and cultured in vitro,which formed embryoid bodies（EBs） after 5 days suspension.The differentiation of EBs was induced by ascorbic acid in various concentrations,which at different times underwent immunocytochemistrical staining for identifying the expression of primordial myocardium-specific transcription factor GATA-4 and cardiac troponin T（cTnT）.Results The maximum cardiac differention rate of hEGC was found at a concentration of 0.1mg/ml ascorbic acid with the differentiatiation rate of （85.2±1.8）% on the 3rd week,which was significantly higher than that in the control.After induction with ascorbic acid,hEGCs shaped fusiform,the cell processes affiliated with each other and the cells formed cell lines on the 4th week.GATA-4 expression appeared to be weakly positive on the 1st week and became the strongest on the 3rd week.The cells began to express cTnT on the 2nd week,which was strongly positive on the 3rd week and became stronger on the 4th week.Conclusion Ascorbic acid could enhance the differentiation of hEGCs into cardiomyocytes,with which we can establish a method of in vitro differentiation of hEGCs towards cardiomyocytes.

关 键 词：抗坏血酸 人胚胎生殖细胞 心肌细胞 

分 类 号：R814[医药卫生—影像医学与核医学]