麝香组分诱导成年大鼠骨髓间质干细胞体外定向分化为神经元样细胞的能力

The Ability of Adult Rat Bone Mesenchymal Stem Cells Differentiating into Neurons-like Cells with Musk's Component in vitro

【目的】探讨麝香组分体外诱导成年大鼠骨髓间质干细胞 (adultratbonemarrowmesenchymalstemcells,rBMM SCs)定向分化为神经元样细胞的能力。【方法】分别采用含麝香多肽或麝香酮的L DMEM培养基体外定向诱导第 5代至 10代的rBMMSCs,并分别应用相差显微镜观察神经元样细胞和免疫细胞组化检测神经元样细胞所表达的神经干细胞特异性标志 (nestin)、神经元特异性标志 [神经元特异性烯醇化酶 (neuronspecificenolase ,NSE)和神经丝蛋白 (neurofilament ,NF) ]及星形胶质细胞特异性标志 [胶质纤维酸性蛋白 (gliafiberacidprotein ,GFAP) ],并进行神经元样细胞记数分析。【结果】成年大鼠BMMSCs在加入含麝香多肽的无血清L DMEM培养基诱导后 ,发现细胞胞体收缩 ,突起伸出 ,形似神经元 ;免疫组化显示诱导出的神经元样细胞NSE、NF、nestin表达阳性 ,GFAP阴性。神经元样细胞记数分析发现 :rBMMSCs经过麝香多肽诱导后 ,NSE+ 神经元样细胞和NF H+ 神经元样细胞的比例分别高达 93 5和 88 2 %;而用含麝香酮的无血清L DMEM培养基却无上述变化。【结论】麝香多肽能在体外诱导rBMMSCs定向分化为神经元样细胞 ,而麝香酮却无此能力。

To investigate the ability of the adult rat bone marrow mesenchymal stem cells (rBMMSCs) differentiate into neurons like cells with Musk's component in vitro rBMMSCs(passage 5 to 10) was pretreated with DMEM/low glucose(L DMEM) containing 20%FCS and 10 μg/L basic fibroblastic growth factor(bFGF) for 24 h,then was induced with induction media consisting of L DMEM and Musk's polypeptide(Mu p) or L DMEM and muscone(Mu n) for 5 h to 7 d The differentiating cells was observed by phase contrast microscopy, and the neural cell markers,including Neuron specific enolase(NSE), neurofilament(NF), Nestin,and glial fibrillary acaidic protein(GFAP), were detected by immunohistochemistry The NSE + and/or NF H + neurons like cells were counted by microscopy One treatment protocols with Mu p,not with Mu n, can induce rBMMSCs differentiated into neuron like cells in the present study The neuron like cells expressed neuron specific marker,such as Nestin,NSE, and NF H,but could not express GFAP, which was the astrocytes specific marker With this differentiation protocol,about 93 5% and 88 2 of the rBMMSCs was NSE positive and NF positive cells, respectively [Conclusion] From these findings,we could conclude that rBMMSCs could be explanded,passaged,and induced to differentiate into neurons like cells with Mu p(not with Mu n) in vitro. Consequently, adult rBMMSCs can also be induced to overcome their mesenchymal commited fates and may constitute an unlimited and accessible cellular reservoir for the treatment of a variety of neurologic diseases