体外诱导人脐带间充质干细胞向神经干细胞的分化

Induced differentiation of human umbilical cord mesenchymal stem cells into neural stem cells in vitro

背景:人脐带间充质干细胞较骨髓、脂肪等来源的间充质干细胞易取材、更为原始,并不受伦理、法律的限制,是中枢神经系统损伤修复最具治疗潜力的细胞来源之一。目的:探讨重组人碱性成纤维生长因子、重组人表皮生长因子定向诱导人脐带间充质干细胞分化为神经干细胞的潜能。方法:采用碱性成纤维生长因子、表皮生长因子作为诱导剂,体外诱导人脐带间充质干细胞;在倒置显微镜下动态观察细胞形态学变化,并用免疫荧光法检测人脐带间充质干细胞向神经干细胞分化的情况,实时荧光定量PCR技术检测诱导前后神经干细胞标记蛋白巢蛋白在mRNA水平的相对表达量。结果与结论:形态学观察经诱导后的人脐带间充质干细胞呈神经干细胞球改变;免疫荧光法及实时荧光定量PCR技术均可检测到神经干细胞标记物巢蛋白,并可进一步分化表达神经元标志蛋白神经元特异性烯醇化酶、微管相关蛋白2以及神经胶质细胞蛋白、神经胶质原纤维酸性蛋白。碱性成纤维生长因子、表皮生长因子可定向诱导人脐带间充质干细胞向神经干细胞及神经元、神经胶质细胞分化。

BACKGROUND：Compared with mesenchymal stem cells from bone marrow and fat, human umbilical cord-derived mesenchymal stem cells, as one of the most potential cellsources to repair the central nervous system, are easy-based, more primitive, and not limited by ethical and legal. OBJECTIVE：To explore differentiation of human umbilical cord-derived mesenchymal stem cells into neural stem cells induced by the recombinant human basic fibroblast growth factor and recombinant human epidermal growth factor. METHODS：The human umbilical cord-derived mesenchymal stem cells were induced by basic fibroblast growth factor and epidermal growth factor in vitro. The cellmorphology was observed by invert microscope. The＆amp;nbsp;differentiation status was detected by immunofluorescence. The Nestin expression in mRNA level before and after induction was detected by real-time PCR. RESULTS AND CONCLUSION：The neural stem cellbal s were observed after induction. And the Nestin was detected by immunofluorescence and real-time PCR. Nestin could further differentiate to the neuronal markproteins neuron-specific enolase, microtubule-associated protein 2 protein and glial fibril ary acidic protein. Results from this study show that basic fibroblast growth factor and epidermal growth factor can induce human umbilical cord-derived mesenchymal stem cells to differentiate to neural stem cells, neurons and glial cells.