共培养方法诱导3种间充质干细胞向神经细胞分化的比较

Comparison of three kinds of mesenchymal stem cells differentiating into nerve cells under co-culture induction

背景:尝试创建更加类似于人体的微环境,从而可以诱导人骨髓间充质干细胞、人胎盘间充质干细胞、人脐血间充质干细胞定向分化。目的:观察与神经细胞共培养诱导人骨髓间充质干细胞、人胎盘间充质干细胞、人脐血间充质干细胞向神经细胞分化的可行性。方法:体外分离培养人骨髓间充质干细胞、人胎盘间充质干细胞、人脐血间充质干细胞,采用Transwell共培养装置建立与神经细胞共培养体系,观察3种细胞的形态变化,在共培养四五天后用免疫荧光染色方法检测3种细胞中神经元特异性烯醇化酶的表达,以单纯低糖DMEM培养基培养的间充质干细胞为对照组。结果与结论:(1)3种间充质干细胞生长伸展,形成突起,并可互相形成连接。神经元特异性烯醇化酶阳性表达率:脐血间充质干细胞>胎盘间充质干细胞>骨髓间充质干细胞;(2)对照组中3种间充质干细胞均没有神经样形态结构,神经元特异性烯醇化酶表达阴性;(3)结果表明,神经细胞提供的共培养微环境对3种间充质干细胞分化为神经元均具有诱导和促进作用。

BACKGROUND： Scholars have been trying to create a microenvironment similar to the human body, which can induce the directional differentiation of mesenchymal stem cells from human bone marrow, placenta and umbilical cord blood. OBJECTIVE： To compare the neuronal differentiation of human bone marrow mesenchymal stem cells, human placental mesenchymal stem cells and human umbilical cord blood mesenchymal stem cells induced by co-culture with nerve cells. METHODS： Human bone marrow mesenchymal stem cells, human placental mesenchymal stem cells and human umbilical cord blood mesenchymal stem cells cultured in vitro were co-cultured with nerve cells using the Transwell system. The morphological changes of three kinds of cells in the co-culture system were detected. After co-culture for4-5 days, immunofluorescence staining was used to measure the expression of neuron-specific enolase in cells. Mesenchymal stem cells only cultured in low glucose DMEM medium were used as controls. RESULTS AND CONCLUSION： These three kinds of mesenchymal stem cells were extended, and interconnected processes were detective. The positive expression of neuron-specific enolase was highest in the human umbilical cord blood mesenchymal stem cells followed by human placental mesenchymal stem cells and human bone marrow mesenchymal stem cells in order. In the control group, none of the three kinds of mesenchymal stem cells have neuronal morphology, and the expression of neuron specific enolase was negative for the immunofluorescence staining. To conclude, microenvironment provided by nerve cells can induce these three kinds of mesenchymal stem cells to differentiate into neurons.