许旺细胞培养液诱导NRG1蛋白转染骨髓基质干细胞向类神经元样细胞的转化

背景：国内外研究人员已经利用化学方法或单纯与许旺细胞共培养方法成功将骨髓基质干细胞诱导为许旺细胞样细胞,但仍存在一些弊端,前者对细胞有一定毒性,后者诱导速度慢,耗时较长。目的：探讨大鼠的许旺细胞培养液诱导NRG1转染的骨髓基质干细胞分化成神经组织细胞的可行性。方法：利用NRG1转染骨髓基质干细胞,待生长状态稳定后,将培养液更换为许旺细胞的培养液,置于37℃、体积分数为5%CO2培养箱中培养。结果与结论：经许旺细胞培养液诱导10 d后,光镜下可观察到NRG1转染的骨髓基质干细胞胞体开始回缩,呈梭形,出现许旺细胞样细胞,呈岛屿状紧密排列,免疫荧光染色鉴定诱导后细胞S-100、NSE和GFAP呈阳性表达,NSE表达阳性率为25.32%,GFAP表达阳性率为38.69%,S-100表达阳性率为35.99%,结果表明许旺细胞分泌的细胞因子和NRG1蛋白共作用可以诱导骨髓基质干细胞高效地向神经组织细胞分化,并表达许旺细胞、神经元和胶质细胞表面抗原。

Schwann cells induce the differentiation of neuregulin-1-transfected bone marrow stromal stem cells into neuron-like cells

Abstract BACKGROUND： Bone marrow stromal stem cells have been successfully induced by chemical method or co-cultured with Schwann cells to differentiate into Schwann cell-like cells, but there are still some drawbacks. The former has some toxicity to cells, and the latter can take a long time for cell induction. OBJECTIVE： To explore the feasibility that Schwann cell medium induces the neuronal differentiation of neuregulin-l-transfected bone marrow stromal stem cells. METHODS： Neuregulin-1 was used to transfected bone marrow stromal cells. When the cells were at stable state the culture medium was replaced by Schwann cell medium, placed at a 37 ℃, 5% CO2 incubator. RESULTS AND CONCLUSION： After 10 days of induced culture in Schwann cell conditioned medium, neuregulin-l-transfected cells were retracted in a fusiform shape under light microscope, and there were Schwann cell-like cells that were closely arranged in a island-like shape. Results from immunofluorescence staining showed that induced bone marrow stromal cells were positive for S-100, neuron-specific enolase and giial fibrillary acidic protein, and the positive rates were 35.99%, 25.32% and 38.69%, respectively. Experimental findings suggest that the Schwann cells-secreted cytokines and neuregulin-1 protein can co-induce the differentiation of bone marrow stromal stem cells efficiently into nerve cells that express the surface antigens of Schwann cells, neurons and glial cells.