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【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。从而可为神经组织工程和临床上各类神经疾病的神经移植及细胞替代治疗提供大量的种子细胞
【Objective】 To investigate the ability of adult rat bone marrow mesenchymal stem cells (rBMM SCs) to differentiate into neuron-like cells in vitro. 【Methods】 rBMMSCs from the fifth passage to the tenth passage were induced in vitro using L DMEM medium containing muscarine polypeptide or muskone, respectively. The neuron-like cells and immunocytochemistry were used to observe the expression of neuron-like cells by phase contrast microscopy. The neural stem cell-specific (nestin), neuron specific markers [neuron specific enolase (NSE) and neurofilament (NF)] and astrocyte-specific markers [collagen] Fibrillary acidic protein (GFAP) and neuronal-like cell count analysis. 【Results】 Adult rat BMMSCs were induced by the addition of serum-free L DMEM medium containing musk polypeptide and the cell bodies were found to have contracted and protruding processes resembling neurons. Immunohistochemistry showed that the induced neurons-like cells NSE, NF, Nestin expression is positive and GFAP is negative. Neuron-like cell count analysis showed that the proportion of NSE+ neuron-like cells and NF H+ neuron-like cells was as high as 93 5 and 88 2%, respectively, after rBMMSCs were induced by muscarine polypeptide; however, they were cultured with serum-free L DMEM containing muconone. However, there is no such change. [Conclusion] Musk polypeptide can induce rBMMSCs to differentiate into neuron-like cells in vitro, but muskone does not have this ability. Thus, a large number of seed cells can be provided for neural transplantation and cell replacement therapy in nerve tissue engineering and clinical neurological diseases.