肝细胞生长因子对鼠胚胎干细胞分化为心肌细胞的作用  

The effect of hepatocyte growth factor on differentiation of murine embryonic stem cells into cardiomyocytes

作　　者：孙晓冬[1] 刘光耀[2] 冯春梅[3] 杨清东[3] 菜克瑞 穆寅东[1] 杨国宏[1] 

机构地区：[1]牡丹江医学院组胚学与胚胎学教研室,黑龙江牡丹江157011 [2]牡丹江医学院附属红旗医院,黑龙江牡丹江157011 [3]牡丹江医学院生物技术实验室,黑龙江牡丹江157011

出　　处：《解剖科学进展》2015年第5期489-492,共4页Progress of Anatomical Sciences

基　　金：黑龙江省教育厅科学技术研究项目(No 12531735)

摘　　要：目的观察肝细胞生长因子(HGF)在胚胎干细胞(ESC)向心肌细胞分化过程中的作用。方法制备小鼠胚胎成纤维细胞饲养层(FL),复苏的E14细胞接种在FL上,经直接悬浮法形成拟胚体(EBs),诱导组每孔加2m L胚体完全培养液再加15 ng/m L HGF,对照组只加入胚体完全培养液每孔2ml,免疫荧光染色激光共聚焦扫描显微镜下观察细胞心肌特异性蛋白Mlc-1v的表达,透射电子显微镜下观察分化细胞的超微结构特征。结果制备的FL细胞为长梭型,呈条索状或漩涡状排列,复苏在FL上的ES-E14细胞呈圆形、椭圆形克隆,经悬浮培养后即可见球形悬浮状EBs。诱导分化后第12d的细胞其心肌Mlc-1v蛋白表达阳性,透射电子显微镜下可见细胞核的周围有许多不规则成束的肌丝分布。结论肝细胞生长因子可以促进胚胎干细胞分化为心肌样细胞。Objective To study the effect of hepatocyte growth factor on differentiation of murine embryonic stem cells（m ES） into cardiomyocytes. Methods The fibroblast feeder layer cells were prepared, on which the embryonic stem（ES） cells were cultured to form the embryoid bodies（EBs） via the direct suspension method. The expression of Mlc-1v protein was examined by immunofluorescence and observed by confocal laser scanning microscopy, the ultrastructural characteristics of m ES cell-derived cardiomyocytes were observed by transmission electron microscopy. Results The positive expression of Mlc-1v protein was found in cardiomyocytes, the irregular actin filaments bundle was observed under electronmicroscope at the 12 th day Conclusion hepatocyte growth factor promotes the differentiation of embryonic stem cells into cardiomyocytes

关 键 词：胚胎干细胞 心肌细胞 肝细胞生长因子 小鼠 

分 类 号：R329.4[医药卫生—人体解剖和组织胚胎学]