体外定向诱导小鼠骨髓间充质干细胞分化为神经元样细胞

The Research of rMSCs Induced and Differentiated into Neuron-like Cells in vitro

[目的]分离和克隆小鼠骨髓间充质干细胞,并诱导分化为神经元样细胞。[方法]利用Percoll密度梯度离心的方法进行小鼠骨髓间充质干细胞分离及培养,分别采用β-巯基乙醇和BHA诱导分化为神经元样细胞。用神经元特异性烯醇化酶和神经纤维丝蛋白免疫细胞化学方法对已分化的神经元样细胞进行鉴定和分化率分析。[结果]经2种诱导剂诱导的小鼠骨髓间充质干细胞均出现神经元样细胞的分化,胞体呈神经元状,伸出较长轴突样和树突样突起且有分支。而且免疫细胞化学鉴定显示,诱导分化后的神经元样细胞神经元特异性烯醇化酶染色和神经纤维丝蛋白免疫细胞化学染色均呈阳性,β-巯基乙醇诱导分化细胞表达NSE的阳性率为(86.8±2.7)%;诱导分化细胞表达Neurofilament200 kD的阳性率为(75.2±2.3)%;BHA诱导分化细胞表达NSE的阳性率为(80.5±2.2)%;诱导分化细胞表达Neurofilament200 kD的阳性率为(73.2±1.6)%。[结论]小鼠骨髓间充质干细胞能够在诱导剂β-巯基乙醇和BNA的诱导下体外诱导分化为神经元样细胞。

[Objective]The research compared whole-marrow isolation with density gradient isolation for the purification of rMSCs,and investigated the influence of different culture mediums and serums in culture rMSCs,together with the ability of differenciation into neuron-like cells of rMSCs.[Method]Density gradient isolation combined with adhenerce isolation can purify rMSCs,which was able to differentiate into neuron-like cells and Adipogenic cells,accordingly it provide powerful technical support for the futher study of rMSCs differentiate into neuron like cells.In the research rMSCs（mice barrow mesenchymal stem cells） had been isolated and purified,together with the ability of differenciation into neuron-like cells of rMSCs during cultured with β-Me and BHA.The rate of neuron-like cells induced by β-ME and BHA expressed NSE and NF had also been researched by using the assays of cellular immunity.[Result]The result proved that mouse bone mesenchymal stem cells appeared the morphologic of neuron-like cells after it was induced by β-ME and BHA.The morphology of the cells like the body of neuron cells which were star-like and came to form long branche.It was proved by immunochemist that the neuron-like cells expressed NSE and NF after induced,the rate of neuron-like cells induced by β-ME expressed NSE was（86.8±2.7）%;and the rate of neuron-like cells induced by β-ME expressed Neurofilament 200 kD was（75.2±2.3）%;the rate of neuron-like cells induced by BHA expressed NSE was（80.5±2.2）%;and the rate of neuron-like cells induced by BHA expressed Neurofilament 200 kD was（73.2±1.6）%.[Conclusion]It was proved that mouse mesenchymal stem cells could be differentiated into neuron-like cells in vitro by β-ME or BHA.