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[目的]研究人脐带Wharton胶间充质干细胞(WJMSCs)的生物学特性及向雪旺细胞诱导分化的可能性,为神经组织工程提供新的种子细胞。[方法]去除新鲜人脐带动、静脉和脐带外膜组织,获得Wharton胶,采用组织块培养法获得脐带Wharton胶中的干细胞,免疫荧光鉴定WJMSCs的细胞表面特异标记CD44,CD105,CD34,CD45,Stro-1,Vimentin和Nestin,评价干细胞的生物学特性;采用b-FGF和Forskolin等诱导WJMSCs向雪旺细胞分化,光镜观察诱导后细胞形态的变化;免疫荧光染色鉴定雪旺细胞特异性标记物p75和GFAP的表达;Western分析诱导前后雪旺细胞特异性标记物GFAP的表达。[结果]分离培养的WJMSCs免疫荧光染色CD44、CD105、Stro-1和Vi-mentin表达阳性,而CD34、CD45和Nestin表达阴性,具有间充质干细胞的生物学特性;WJMSCs经过胶质细胞生长因子的作用,光镜下发现诱导的细胞形态与雪旺细胞相似;免疫荧光染色p75和GFAP阳性;Western结果显示诱导的雪旺细胞标记物GFAP表达阳性。[结论]人脐带Wharton胶间充质干细胞可诱导分化成为雪旺样细胞,其表型和分子特征与雪旺细胞相似,诱导分化的人脐带Wharton胶间充质干细胞是一种理想的神经组织工程的种子细胞。
[Objective] To investigate the biological characteristics of human umbilical cord Wharton glue-derived mesenchymal stem cells (WJMSCs) and the possibility of inducing differentiation into Schwann cells, and to provide new seed cells for neural tissue engineering. [Method] Wharton ’s gum was obtained from the umbilical cord, umbilical cord and umbilical cord of fresh human umbilical cord. Wharton’ s glue was obtained from the umbilical cord by tissue block culture. The surface markers CD44, CD105, CD34 and CD45 of WJMSCs were identified by immunofluorescence. Stro-1, Vimentin and Nestin were used to evaluate the biological characteristics of stem cells. WJMSCs were induced to differentiate into Schwann cells by b-FGF and Forskolin. The changes of cell morphology were observed under light microscope. Schwann cell specific markers were identified by immunofluorescence staining The expressions of p75 and GFAP were detected by Western blot. The expression of GFAP was detected by Western blot. [Results] The immunofluorescent staining of isolated WJMSCs showed positive expression of CD44, CD105, Stro-1 and Vi-mentin, but negative expression of CD34, CD45 and Nestin, which had the biological characteristics of mesenchymal stem cells. WJMSCs were transfected with glial cell growth factor The results showed that the induced cell morphology was similar to that of Schwann cells under the light microscope. Immunofluorescence staining showed that p75 and GFAP were positive. Western blot showed that the induced GFAP expression was positive in Schwann cells. [Conclusion] The human umbilical cord Wharton glue-derived mesenchymal stem cells can differentiate into Schwann-like cells with similar phenotype and molecular characteristics as Schwann cells. The induced differentiation of human umbilical cord Wharton’s mesenchymal stem cells is an ideal nerve tissue Engineering seed cells.