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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、14d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。
OBJECTIVE: To investigate the potential of rat bone marrow mesenchymal stem cells (BMSCs) to differentiate into neurons in vitro and the apoptosis after differentiation. Methods: Bone marrow mesenchymal stem cells were cultured in vitro. The third generation of MSCs were induced to differentiate by induction medium containing epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF), and the differentiated cells were differentiated Time 3,7,14 d into A, B, C three groups. Three groups of differentiated cells were subjected to NSE immunohistochemistry, MAP2 immunofluorescence and TUNEL apoptotic cells. Results: The cells in groups A, B and C were similar in shape to neurons in morphology and expressed NSE and MAP2, but the expression of cells in group C was weaker than that in group B, and the percentage of positive cells was less than that in group B (P <0.05). The proportion of apoptotic cells in group A, B and C increased gradually (P <0.05). CONCLUSION: Induction medium containing epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) can induce rat bone marrow mesenchymal stem cells to differentiate into neurons. With the induction of time, the differentiated cells decreased, Apoptosis increased.