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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。
Objective To analyze the changes of protein expression profile during the differentiation of human embryonic germ cells (hEGC) into cardiomyocytes. Methods hEGC was induced by tissue culture in vitro. The hEGC was induced to differentiate into cardiomyocytes by using ascorbic acid. The expression of cT-nT was detected by immunofluorescence. The cells were harvested and cultured for 2 weeks. Total proteins were identified by iTRAQ labeling, total protein by liquid chromatography (LC), and differential proteins by tandem mass spectrometry (MS / MS). Results The positive cells were cTnT after induction. A total of 349 proteins were identified by mass spectrometry and database search, of which 27 proteins were significantly different. Conclusion hEGC can differentiate into cardiomyocytes; proteomics method can screen high-throughput screening of important proteins related to hEGC differentiation into cardiomyocytes.