摘要
目的探讨大鼠骨髓间质干细胞(mesenchymalstemcells,MSCs)体外定向分化为心肌样细胞。方法采用骨髓细胞体外培养技术分离大鼠骨髓MSCs,用流式细胞仪检测细胞表面标志;用第二代骨髓MSCs进行体外心肌细胞诱导分化;采用免疫组化、透射电镜及RTPCR鉴定心肌细胞。结果大鼠骨髓MSCs细胞表面抗原CD29、CD44阳性,CD15、CD33、CD34、HLADR阴性;经5氮胞苷体外诱导向心肌细胞分化,其细胞免疫组化呈结蛋白、肌动蛋白和肌球蛋白重链强阳性;在透射电镜下,诱导后细胞具有条索状肌丝结构;RTPCR显示诱导组细胞后转录结蛋白、肌动蛋白和肌球蛋白重链mRNA水平增加。结论大鼠骨髓MSCs体外在5氮胞苷作用下可定向分化为心肌样细胞,为应用MSCs治疗心肌损伤提供了动物实验依据。
Objective: To investigate rat bone marrow mesenchymal stem cells (MSCs) differentiated into cardiomyocyte phenotype in vitro. Methods: Rat MSCs were isolated from adult bone marrow. Surface antigentic features of the MSCs were analyzed by flow cytometry. The MSCs from passage 2 were selected for differentiating to cardiomyocytes. The myogenic cells differentiated were evaluated by immunohistochemistry stained for myosin heavy chain (MHC), desmin and cardiac actin, transmission eletron microscope and RT-PCR. Results: The MSCs were positive for CD29、CD44 and negative for CD15、CD33、CD34、HLA-DR. The MSCs differentiated, cardiomyocytes has been shown positive for myosin heavy chain (MHC), desmin and cardiac actin. The myofilament like apperance can be found in the cardiomyocytes by transmission eletron microscope. The mRNA of myosin heavy chain (MHC), desmin and cardiac actin was highly expressed using RT-PCR. Conclusion: The results showed rat MSCs can be differentiated to cardiomyocyte phenotype in vitro after 5-azacytidine treatment. It can be served as optimal stem cells source for repairing myocardium tissue impairment. [
出处
《江苏大学学报（医学版）》
CAS
2004年第416期461-464,共4页
Journal of Jiangsu University：Medicine Edition
基金
国家自然科学基金资助项目(30471938)
江苏省自然科学基金资助项目(BK2002006)
江苏自然科学基金重点资助项目(BK2002208)
