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  

The study on neural induction of mesenchymal stem cells in vitro and apoptosis of the neuron-like cells

作　　者：程峰[1] 郝怀勇[1] 田和平[1] 代学良[1] 李立新[1] 

机构地区：[1]南京医科大学第一附属医院神经外科,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2010年第1期54-58,共5页Journal of Nanjing Medical University(Natural Sciences)

基　　金：江苏省"兴卫工程"医学重点人才基金资助项目(RC2007062)

摘　　要：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 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.

关 键 词：骨髓间充质干细胞 细胞分化 神经样细胞 凋亡 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学]