人胚胎生殖细胞向心肌细胞诱导分化及质谱分析

目的分析人胚胎生殖细胞（hEGC）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hEGC,采用抗坏血酸诱导hEGC向心肌细胞分化,通过免疫荧光法检测诱导后细胞中肌钙蛋白T（cT-nT）的表达;分别提取hEGC及诱导分化2周后的细胞总蛋白,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 hEGC能诱导分化为心肌细胞;蛋白质组学方法可以高通量筛选hEGC向心肌细胞分化相关的重要蛋白。

Mass Spectrum Analysis of Human Embryonic Germ Cell Induced to Differentiate into Cardiomyocytes

Objective To analyze the difference of protein profiles between human embryonic germ cell（hEGC） and cardiomyocytes differentiated from hEGC.Methods Tissue was cultured in vitro to obtain hEGC,differentiation of hEGC induced to cardiocytes by ascorbic acid,cTnT detected by immunofluo-rescence on differentiated cells and cell total proteins of the hEGC and cardiomyocytes differentiated from hEGC were extracted over a period of 2 week and labeled by isobaric tags for relative and absolute quantitation（iTRAQ）.The labeled protein was isolated by liquid chromatography（LC） for identifying the different expressing of protein by MS/MS.Results cTnT expression was positive in differentiated cells;349 kinds of protein were identified by MS analysis and database search,among which 27 kinds of protein were differently expressed.Concusion hEGC is induced to differentiated into cardiomyocytes;proteomic approach can screen the important proteins in a high throughput which is related to differentiation of hEGC induced to cardiomyocytes.