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背景:与骨髓间充质干细胞相比,脐带血中的细胞被视为“非常年轻”的细胞,其增殖和分化能力不会随着捐助者的年龄增加而减低,可能是一个极好的骨髓间充质干细胞的替代来源。目的:观察体外诱导脐血间充质干细胞分化成心肌细胞的特点。设计、时间及地点:观察性实验,于2005-03/2007-02在北京世纪坛医院完成。材料:收集获知情同意健康产妇脐血细胞。方法:分离单个核细胞,从中进一步分离间充质干细胞,传代培养至第3代,应用免疫荧光流式细胞仪标记间充质干细胞特异性抗原CD34,CD44和CD90。5-氮胞苷诱导分化4周后,免疫组织化学染色和反转录-聚合酶链反应分别检测心肌细胞标志物肌钙蛋白Ⅰ,GATA4和β-肌球蛋白重链的表达。主要观察指标:心肌细胞标志物肌钙蛋白Ⅰ,GATA4和β-肌球蛋白重链的表达。结果:脐血源性间充质干细胞经5-氮胞苷诱导分化后,呈现成纤维细胞样形态和克隆增殖特点。免疫分型与骨髓来源间质干细胞一致,且免疫组织化学染色和反转录-聚合酶链反可检测到肌钙蛋白Ⅰ,GATA4和β-肌球蛋白重链的表达。结论:脐血源性间充质干细胞能够被诱导分化成心肌样细胞,并显示为心肌细胞的表观特征。
Background: Cells in umbilical cord blood are considered “very young” cells compared to bone marrow-derived mesenchymal stem cells and their ability to proliferate and differentiate may not decrease as the donor’s age increases and may be an excellent Of alternative sources of bone marrow mesenchymal stem cells. Objective: To observe the characteristics of differentiating umbilical cord blood mesenchymal stem cells into cardiomyocytes in vitro. DESIGN, TIME AND SETTING: Observational experiments were performed at Beijing Millennium Monument Hospital from March 2005 to February 2007. MATERIALS: Collection of informed cordial healthy umbilical cord blood cells. Methods: Mononuclear cells were isolated and mesenchymal stem cells were further isolated and passaged to the third passage. Induction of differentiation of mesenchymal stem cell-specific antigen CD34, CD44 and CD90.5-azacytidine by immunofluorescence flow cytometry Four weeks later, the expression of cardiac troponin I, GATA4 and beta-myosin heavy chain were detected by immunohistochemical staining and reverse transcription-polymerase chain reaction. MAIN OUTCOME MEASURES: Cardiomyocyte markers troponin I, GATA4 and β-myosin heavy chain expression. Results: After umbilical cord blood-derived mesenchymal stem cells differentiated by 5-azacytidine, they showed the characteristics of fibroblast-like morphology and clonal proliferation. Immunophenotyping was consistent with bone marrow-derived mesenchymal stem cells, and the expression of troponin I, GATA4 and β-myosin heavy chains was detected by immunohistochemistry and reverse transcription-polymerase chain reaction. CONCLUSION: Umbilical cord blood-derived mesenchymal stem cells can be induced to differentiate into cardiomyocyte-like cells and show the apparent characteristics of cardiomyocytes.