人脐带Wharton胶间充质干细胞的生物学特性及雪旺细胞分化研究

Biological characteristics of human umbilical cord Wharton’s jelly-derived mesenchymal stem cells and their differentiation into Schwann-like cells

[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 Wharton＇ s jelly -derived mesenchymal stem cells （WJMSCs） and their differentiation into Schwann cell -like cells, and to provide a new cell seed source for nerve tissue engineering or cell therapy. [ Methods] Umbilical arteries, veins and umbilical cord tunica externa were removed , and the remaining tissue （Wharton＇ s jelly） was harvested. The umbilical cord was cut into small fragments and cultured with DMEM in orderto gather. Cultured WJMSCs were confirmed by the detection of MSC - specific cell - surface markers. WJMSCs were differentiated along a glial cell lineage using an established cocktail of growth factors. Immunocytochemical staining and Western blot were used to evaluate the characteristics of differentiated WJMSCs. [ Results] WJMSCs were immunonegative for the haemopoietic cell- surface markers （CD34 and CD45） and neural stem cell marker （nestin） , and immunopositive for MSC surface markers CD44, CD105, Stro - 1 and vimentin. WJMSCs treated with a mixture of glial growth factors adopted a spindle - like morphology similar to Schwann cells, lmmunocytocbemical staining and Western blot analysis revealed that the treated cells expressed the glial markers p75 and glial fibrillary acidic protein indicative of differentiation. [ Conclusion] WJMSCs can be differentiated into the cells that are Schwann - like in terms of morphologic features and phenotype and may be suitable as Schwann - cell substitutes for nerve repair in clinical application.