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目的:骨髓间质干细胞体外诱导和非诱导培养向心肌细胞分化的结果的对比。方法:实验于2003-03/2004-12在阜外心血管病医院中心实验室完成。①分离犬骨髓间质干细胞。②于体外培养,应用不同浓度(0,6,8,10,12,14,20μmol/L)的5-氮胞苷定向诱导并连续传代培养4周,未诱导的细胞培养8周。③进行细胞形态学、细胞免疫组织化学、透射电镜鉴定。结果:①骨髓间质干细胞于5-氮胞苷诱导培养2周时,α-肌动蛋白、TroponinI染色阴性;4周时,α-肌动蛋白染色阳性,TroponinI阴性;电镜下可见肌丝结构,其中8mmol/L5-氮胞苷诱导的间质干细胞肌丝结构排列较规则。②骨髓间质干细胞非诱导培养8周时,α-肌动蛋白染色阳性,TroponinI阴性,电镜下亦可见肌丝结构形成;而培养4周时全部为阴性。结论:骨髓间质干细胞经5-氮胞苷诱导培养4周及未诱导连续培养8周均可定向转化为具有肌丝结构的肌样细胞,而非心肌样细胞。
OBJECTIVE: To compare the results of differentiation of cardiomyocytes induced by in vitro and non-induced culture of bone marrow mesenchymal stem cells. Methods: The experiment was performed in Fuwai Cardiovascular Hospital Central Laboratory from March 2003 to December 2004. ① isolated canine bone marrow mesenchymal stem cells. ② In vitro culture, 5-azacytidine with different concentrations (0, 6, 8, 10, 12, 14 and 20 μmol / L) was used for directional induction and continuous subculture for 4 weeks, and uninduced cells were cultured for 8 weeks. ③ cell morphology, cellular immunohistochemistry, transmission electron microscopy. RESULTS: ①BMSCs were negative for α-actin and Troponin I staining after induction of 5-azacytidine for 2 weeks. Α-actin staining and Troponin I were negative for 4 weeks. The myofilament structure , Of which 8mmol / L 5-azacytidine-induced mesenchymal stem cells arranged more regular filamentous structure. ② After 8 weeks of culture, bone marrow mesenchymal stem cells were positive for α-actin staining and negative for TroponinI, and myofilament structure was also observed under electron microscope. All the cells were negative for 4 weeks. CONCLUSION: Bone marrow mesenchymal stem cells can be directionally transformed into myofibroblasts with myofilament structure rather than cardiomyocytes after 4-week induction with 5-azacytidine and 8-week without induction.