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目的:探讨骨髓间质干细胞(MSCs)体外诱导分化为心肌细胞的可行性,为心肌梗死治疗提供理想的细胞材料。方法:分离大鼠下肢骨获取MSCs进行培养;5-氮胞苷(5-aza)诱导24h后继续培养;免疫细胞化学检测细胞对连接蛋白-43和α横纹肌肌动蛋白的表达;逆转录-聚合酶链反应(RT-PCR)进一步了解心肌细胞特异性蛋白的表达。结果:MSCs经5-aza诱导后,细胞形态不规则;诱导后3周10μmol/L 5-aza组细胞连接蛋白-43、α横纹肌肌动蛋白表达阳性。RT-PCR显示10μmol/L 5-aza诱导后3周的细胞可表达心肌肌钙蛋白I、α心肌肌动蛋白。结论:MSCs体外经5-aza诱导后可分化为心肌细胞。
Objective: To investigate the feasibility of differentiation of bone marrow mesenchymal stem cells (MSCs) into cardiomyocytes in vitro and to provide ideal cell material for the treatment of myocardial infarction. Methods: MSCs were isolated from rat lower extremity bone for culturing. 5-azacytidine (5-aza) was cultured for 24 hours and then cultured. Immunocytochemistry was used to detect the expression of connexin-43 and α-striated actin in cells. Reverse transcription- Polymerase chain reaction (RT-PCR) further understanding of cardiomyocyte-specific protein expression. Results: The morphology of MSCs induced by 5-aza was irregular. The expression of connexin-43 and α-striated actin in 10μmol / L 5-aza group was positive at 3 weeks after induction. RT-PCR showed that cells at 3 weeks after induction with 10μmol / L 5-aza could express cardiac troponin I and α-actin. Conclusion: MSCs can differentiate into cardiomyocytes after induced by 5-aza in vitro.