论文部分内容阅读
目的 定向诱导人骨髓间质干细胞(hMSC)分化为神经元样细胞,并检测诱导前后细胞的运动神经元生存(SMN)基因mRNA及蛋白质的表达。方法 分离和纯化hMSC;在体外用碱性成纤维细胞生长子预诱导hMSC,再用二甲基亚砜和丁化羟基苯甲醚诱导hMSC分化为神经元样细胞;用免疫细胞化学方法鉴定诱导前后的细胞是否表达神经元特异性标志物神经元稀醇化酶(NSE)、神经丝蛋白(NF)以及SMN蛋白;用RT- PCR检测诱导前hMSC及诱导后神经元样细胞SMN基因mRNA的表达。结果 hMSC诱导3h后,大多数细胞转变为类似于神经元的形态,有长的突起,相互之间连接呈网状。NSE、NF进行鉴定,结果显示NSE、NF呈阳性。诱导前的hMSC及诱导后的神经元样细胞均可见SMN基因mRNA表达,但诱导后表达增加,差异有统计学意义(P<0 .01),且诱导后神经元样细胞SMN蛋白表达为阳性。结论 hMSC能分化为神经元样细胞,并表达神经元的特异性标志物。hMSC分化的神经元样细胞既能高表达SMN基因的mRNA,也表达SMN蛋白。
Objective To induce the differentiation of human bone marrow mesenchymal stem cells (hMSCs) into neuron-like cells and to detect the expression of motor neuron survival (SMN) mRNA and protein before and after induction. Methods hMSCs were isolated and purified. HMSCs were pre-induced by basic fibroblast growth factor (hMSC) in vitro and then differentiated into neuron-like cells using dimethyl sulfoxide and butylated hydroxyanisole. Neuronal-like cells were induced by immunocytochemistry Before and after the cells expressed neuron-specific marker neuronal alcoholase (NSE), neurofilament protein (NF) and SMN protein; RT-PCR detection of hMSC induction and after induction of neuron-like cells SMN gene mRNA expression . Results After 3 h induction of hMSC, most of the cells turned into neuron-like morphology with long protuberances and reticular connections. NSE, NF were identified, the results showed NSE, NF was positive. SMN gene mRNA expression was observed in both pre-induction hMSC and post-induction neuron-like cells, but the expression was increased after induction (P <0.01), and the expression of SMN protein in neuron-like cells was positive . Conclusion hMSC can differentiate into neuron-like cells and express neurons specific markers. hMSC differentiated neuron-like cells both express SMN mRNA and express SMN protein.