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目的 探索在体外诱导人骨髓间充质干细胞(hMSC)向心肌细胞分化的方法。方法 以5 杂氮胞苷( 5 Aza)对hMSC诱导,利用结蛋白抗体、房性利钠肽抗体以及闰盘蛋白抗体进行免疫组化染色,并进行超薄切片透射电镜观察。结果 经反复多次传代后,细胞形态及抗原表达无改变,始终保持未分化状态和稳定扩增。利用5 Aza对其诱导后第2周始,免疫组化染色发现细胞结蛋白、闰盘蛋白以及房性利钠肽表达均为阳性,不同浓度5 Aza诱导组之间差异无显著性。电镜观察发现这些细胞具有心肌细胞的特点。结论 在体外利用5 Aza可以诱导hMSC向心肌细胞分化。
Objective To explore the method of inducing human bone marrow mesenchymal stem cells (hMSCs) to differentiate into cardiomyocytes in vitro. Methods The hMSCs induced by 5-azacytidine (5 Aza) were used for immunohistochemical staining using antibodies against desmokinin, atrial natriuretic peptide and anti-intercalated proteins, and observed by ultrathin section transmission electron microscopy. Results After repeated passages, the cell morphology and antigen expression did not change, and remained undifferentiated state and stable amplification. At the beginning of the second week after the induction by 5 Aza, the expression of protein was detected by immunohistochemistry, and the protein expression of atrial natriuretic peptide was positive. There was no significant difference between the 5 Aza groups. Electron microscopy found that these cells have the characteristics of myocardial cells. Conclusion In vitro use of 5 Aza can induce hMSC to differentiate into cardiomyocytes.