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目的:研究人脐带血间充质干细胞体外诱导分化为心肌细胞的可行性。方法:由人脐静脉获得间充质干细胞,纯化培养后用不同浓度的5-氮杂胞苷(5-azacytidine)诱导,透射电镜、免疫组化及RT-PCR鉴定诱导后细胞。结果:经5-氮杂胞苷诱导后的脐带血间充质干细胞在透射电镜下观察到细胞内有明显的肌丝,横纹样结构形成,核呈卵圆形,位于细胞中央位置,胞质中可见富集的糖原颗粒及线粒体结构。免疫组化可见肌钙蛋白Ⅰ染色阳性。RT-PCR方法显示能表达缝隙连接膜通道蛋白Connexin-43基因。结论:人脐带血间充质干细胞可在体外经5-氮杂胞苷诱导分化为具有典型结构的心肌细胞。
Objective: To study the feasibility of human umbilical cord blood-derived mesenchymal stem cells differentiating into cardiomyocytes in vitro. Methods: Mesenchymal stem cells were obtained from human umbilical vein. After being purified, the cells were induced with different concentrations of 5-azacytidine, and the cells were identified by transmission electron microscopy, immunohistochemistry and RT-PCR. Results: Umbilical cord blood-derived mesenchymal stem cells induced by 5-azacytidine showed obvious myofilament and transverse pattern formation under transmission electron microscope. The nucleus was oval in shape, located in the central part of the cell. The cytoplasm In the visible enrichment of glycogen particles and mitochondrial structure. Immunohistochemistry showed that troponin I staining was positive. RT-PCR showed that the connexin-43, a connective membrane protein, can be expressed. Conclusion: Human umbilical cord blood-derived mesenchymal stem cells can differentiate into cardiomyocytes with typical structure in vitro by 5-azacytidine.