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目的 :探讨成年犬骨髓基质干细胞 (MSSCs)体外诱导分化为肌源性细胞的可行性。方法 :用贴壁法体外分离MSSCs ,实验组 5 氮胞苷诱导 ,对照组加入等量的培养基 ,用相差显微镜、免疫组织化学和电镜方法观察、鉴定培养细胞。结果 :实验组培养细胞诱导后 4周 ,多数培养细胞由梭形变为杆状 ,免疫组织化学检查结蛋白、平滑肌肌动蛋白和肌钙蛋白Ⅰ染色阳性 ;对照组多数培养细胞为梭形 ,免疫组织化学检查结蛋白染色弱阳性 ,平滑肌肌动蛋白染色弱阳性 ,肌钙蛋白Ⅰ染色阴性 ;电镜下实验组培养细胞内可见肌丝样结构 ,对照组培养细胞无明显肌丝样结构。结论 :成年犬MSSCs体外可诱导分化为肌源性细胞。
Objective: To investigate the feasibility of adult canine bone marrow stromal cells (MSSCs) differentiated into myogenic cells in vitro. Methods: MSCs were isolated by adherent method in vitro. The experimental group was induced with 5-azacytidine, and the control group was treated with the same amount of medium. The cultured cells were identified by phase-contrast microscope, immunohistochemistry and electron microscopy. RESULTS: Four weeks after induction, most of cultured cells changed from spindle to rod shape, immunohistochemical staining of desmin, smooth muscle actin and troponin I. Most of cultured cells in control group were spindle-shaped, immunized Histochemical staining was weakly positive for knot protein, weakly positive for smooth muscle actin staining and negative for troponin I staining. Myofilament-like structure was observed in the cultured cells in the experimental group under electron microscope, and no myofibrillar structure was found in the cultured cells in the control group. Conclusion: Adult canine MSSCs can differentiate into myogenic cells in vitro.