摘要
目的研究青-链霉素(双抗)对小鼠胚胎干细胞(mouse embryonic stem cells,mESCs)向心肌细胞分化的影响。方法采用常规悬滴法培养mESC形成拟胚体(embryonic body,EB),在分化培养基中加入不同浓度的双抗(1.0%、5.0%和10.0%)培养贴壁的EB,选择不同的时间点分化的细胞通过流式细胞术(FCM)、细胞免疫荧光、westem blotting等分析心肌特异性蛋白表达水平的差异。结果小鼠多能干细胞表面标志物SSEA-1表达呈阳性(绿色),H.E染色显示核质比>>1。正常自分化细胞免疫荧光显示cTnT、‘MLC2阳性(绿色),肌节清晰可见。高浓度双抗组,cTnT+阳性细胞率高于对照组和低浓度(1.0%)双抗组,western blotting显示其心肌特异性蛋白cTnT、cTnI、MLC2、connexin43等表达增加(p<0.05)。结论高浓度双抗促进mESCs向心肌细胞分化,且有浓度依赖性。
Objective To explore the role of penicillin-streptomycin on mouse embryonic stem cells differentiation into cardiomyocytes. Methods Firstly, embryonic bodys though hanging drop method, were incubated with penicillin-streptomycin at different concentrations （1.0%, 5.0% and 10.0%）, and then, the number of cardiomyocytes and expression levels of myocardial marker proteins were detected by flow cytometry （FCM）, cellular immunofluorescence, and western blotting, respectively. Results We found that the SSEA-1 of mESC presented green fluorescence, and H.E staining showed karyoplasmic ratio 〉〉1. Besides, cTnT and MLC2 were clearly visible by cellular immunofluorescence, as well as the unique structure of the muscle fibers. Furthermore, the cTnT positive cells rate of 10x group was remarkably higher than that in control group and 1 ~ group, as well as the expressions of cTnT, cTnI, MLC2 and connexin43 （p〈0.05）. Conclusions The high-concentration penicillin-streptomycin promotes the differentiation of mESC into eardiomyocytes in a concentration- dependent manner.
作者
宋海涛
陈静
孙立辉
凌秋平
贾阿娜
张洪亮
SONG Hai-tao CHEN ling LING Qiu-ping SU Hai-xia JIA A-na ZHANG Hong-liang(The second people＇s Hospital of Longgang District, Shenzhen518000, China.)
出处
《中国分子心脏病学杂志》
CAS
2016年第5期1824-1827,共4页
Molecular Cardiology of China
基金
黑龙江省自然科学基金项目(H201364)
黑龙江省卫生厅项目(2011-355)
关键词
小鼠胚胎干细胞
青-链霉素
心肌细胞
分化
Mouse Embryonic Stem Cell
Penicillin-Streptomycin
Cardiomyocyte
Differentiation
