体外诱导骨髓间充质干细胞向神经细胞分化及凋亡的研究
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江苏省“兴卫工程”医学重点人才基金资助项目(RC2007062)


The study on neural induction of mesenchymal stem cells in vitro and apoptosis of the neuron-like cells
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    目的:探讨大鼠骨髓间充质干细胞体外向神经细胞诱导分化的潜能,及分化后的凋亡情况-方法:体外培养大鼠骨髓间充质干细胞,用含有表皮生长因子(EGF),碱性成纤维细胞生长因子(bFGF)的诱导培养基对第三代MSCs进行诱导分化,诱导分化的细胞按分化时间3-7-14 d分成A-B-C三组-三组诱导分化细胞分别进行NSE免疫组织化学,MAP2免疫荧光化学以及TUNEL凋亡细胞检测-结果:A-B-C三组细胞在细胞形态上与神经细胞相似,并且表达神经细胞特异性标记抗原NSE-MAP2,但是C组细胞表达强度弱于B组,并且阳性细胞比例少于B组(P < 0.05)-A-B-C三组TUNEL凋亡阳性细胞比例呈逐渐增多趋势(P < 0.05)-结论:含有表皮生长因子(EGF),碱性成纤维细胞生长因子(bFGF)的诱导培养基可以诱导大鼠骨髓间充质干细胞向神经细胞分化,随着诱导时间的推移,分化细胞状态下降,凋亡比例增多-

    Abstract:

    Objective:To evaluate the neural differentiation potential of rat bone marrow mesenchymal stem cells and apoptosis of the neuron-like cells in vitro. Methods:MSCs were isolated from rat bone marrow,seeded at passage 3 and cultured in L-DMEM with neural cell inductors(EGF,bFGF). Cells were divided three groups according to the time of induction:group A was induced for 3 days;and group B was inducd for 7 days;and group C was induced for 14 days . The markers of neural cells such as NSE,Map2 were detected by immunocytochemistry. TUNEL method was used to calculate the percentages of neuron-like cells apoptosis. Results:MSCs appeared typical morphological neuronal traits after induction,NSE,MAP2 were expressed,but the expression intensity and the percentage of the positive cells attenuated in group C compared with those in group B(P < 0.05). The percentage of apoptosis cells detected by tunnel gradually increased from A to C groups(P < 0.05). Conclusion:Combination of bFGF and EGF can induce MSCs to differentiate into neuron-like cells,but the status of induced cells plummeted and the percentage of apoptosis cells increased in a time-dependent manner.

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程〓峰,郝怀勇,田和平,代学良,李立新.体外诱导骨髓间充质干细胞向神经细胞分化及凋亡的研究[J].南京医科大学学报(自然科学版),2010,(1):54-58

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  • 收稿日期:2009-06-19
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