胚胎生开殖细胞.docVIP

人胚胎生殖细胞向心肌细胞诱导分化及质谱分析的研究 王士珍, 朱旻, 冯琦, 陈永珍 （苏州大学医学部人体解剖与组织胚胎学系，江苏苏州215123） 摘要:目的 分析人胚胎生殖细胞(hEGCs)向心肌细胞分化过程中蛋白质表达谱的改变。方法 组织块培养法体外培养人EG细胞，采用抗坏血酸诱导人EG细胞向心肌细胞分化，通过免疫荧光法检测诱导后细胞中心肌肌钙蛋白T(cTnT)的表达；分别提取人EG细胞及诱导分化2周后细胞蛋白质，采用iTRAQ标记、液相色谱(LC)分离总蛋白，串联质谱(MS/MS)鉴定差异蛋白。结果 诱导后细胞阳性表达cTnT；通过质谱分析和数据库搜索共鉴定出349种蛋白，其中27种蛋白差异显著。结论 人EG细胞能诱导分化为心肌细胞；蛋白质组学方法可高通量筛选人EG细胞向心肌细胞分化相关的重要蛋白。 关键词：人胚胎生殖细胞；心肌肌钙蛋白T；质谱 Mass Spectrum Analysis of Human Embryonic Germ Cells Induced to Differentiate into Cardiomyocytes WANG Shi-zhen, ZHU Min, FENG Qi, CHEN Yong-zhen (Department of Human Anatomy and Histoembryology，Medical School of Soochow University，Suzhou 215123，China) Abstract：Objective To analyze the difference of protein profiles between hu- man embryonic germ cells(hEGCs) and cardiomyocytes differentiated from hEGCs. Methods Tissue was cultured in vitro to obtain hEGCs, and then which were induced to differentiate into cardiomyocytes by Ascorbic Acid. The expression of cardiac Troponin T (cTnT) is detected by Immunofluorescence on differentiated cells; Cell total proteins of the hEGCs and hEGCs induced to differentiate for 2 weeks were extracted respectively and labeled by isobaric tags for relative and absolute quantitation (iTRAQ).The labeled proteins were isolated by liquid chromatography 基金项目：苏州大学医学发展基金资助项目(EE134710) 收稿日期：2011-03-17 作者简介：王士珍(1980-)，男，山东菏泽人，理学硕士，研究方向为人胚胎生殖细胞。通讯作者：陈永珍 (LC), and then the different expressing of proteins were identified by Tandem mass spectrometry (MS/MS) in hEGCs and differentiated cells. Results cTnT was positive expression in differentiated cells;349 proteins were identified by MS analysis and Database search, among which 27 proteins were different expression significantly. Concusions hEGCs are induced to differentiated into cardiomyocytes; Proteomic approach can screen the important proteins in a high throughput which are related to differentiation of hEGCs induced to cardiomyocytes. Key words: human embryonic germ cells; cTnT; Mass Spectrum 人胚胎生殖细胞