全反式视黄酸诱导兔骨髓间充质干细胞向神经细胞的分化

背景：目前,在骨髓间充质干细胞体外向神经细胞分化的实验中,较多采用抗氧化剂法和细胞因子法,但诱导效率低。目的：探讨全反式视黄酸在兔骨髓间充质干细胞向神经细胞分化中的作用。方法：体外分离培养兔骨髓间充质干细胞,取第4代骨髓间充质干细胞观察细胞形态,实验组用0.4μmol/L全反式视黄酸预诱导24h后,改用神经细胞培养基继续培养。神经细胞培养基培养4,8,16及24h,免疫组织化学检测巢蛋白、神经元特异性烯醇化酶和胶质纤维酸性蛋白的表达情况,采用RT-PCR的方法检测巢蛋白和神经元特异性烯醇化酶的表达水平。结果与结论：骨髓间充质干细胞经培养、传代后,细胞贴壁生长,呈长梭形。免疫组织化学检测结果显示：全反式视黄酸诱导组巢蛋白及神经元特异性烯醇化酶呈阳性,诱导后细胞的活力良好。RT-PCR结果显示全反式视黄酸诱导组巢蛋白在诱导前后均有表达,神经元特异性烯醇化酶在诱导后16h可见明显的扩增条带,24h后更加明显。提示全反式视黄酸能够在体外促进兔骨髓间充质干细胞向神经细胞分化。

All-trans-retinoic acid induces the differentiation of rabbit bone marrow mesenchymal stem cells into neural cells

BACKGROUND：At present,antioxidant and cytokine method are used in the study of the differentiation of bone marrow mesenchymal stem cells（BMSCs） into nerve cells in vitro,but the induced efficiency was low.OBJECTIVE：To investigate the role of all-trans-retinoic acid（ATRA） in the differentiation of rabbit BMSCs into neural cells.METHODS：Rabbits BMSCs were isolated and expanded in vitro,and the 4th generations were obtained for observation of cell morphology.After 24 hours of induction using 0.4 μmol/L ATRA,neuronal medium was used in the experimental group.At 4,8,16 and 24 hours following culture in the neuronal medium,immunohistochemistry was used to detect expressions of nestin,neuron specific enolase（NSE） and glial fibrillary acidic protein（GFAP）.RT-PCR was utilized to determine expression of nestin and NSE.RESULTS AND CONCLUSION：Following culture and subculture,BMSCs were adherent and fibroblast-like.Immunohistochemical method demonstrated that ATRA group was positive for nestin and NSE.Following induction,cell viability was good.RT-PCR results showed that ATRA group expressed nestin before and after induction.At 16 hours following induction,NSE exhibited significant amplification band,and became more obvious at 24 hours.These suggest that ATRA could promote the differentiation of rabbit BMSCs into neural cells in vitro.