黄芪诱导人脐带间充质干细胞分化为神经样细胞的实验研究

The research about astragalus mongholicus induce human umbilical cord derived mesenchymal stem cells into neurons

目的 研究原代人脐带间充质干细胞（UCMSCs）的生物学特性及其体外向神经细胞诱导分化的条件,为组织化人工神经的制备提供干细胞资源.方法 无菌条件下从人脐带华尔通胶（Wharton jelly）分离并贴壁培养人UCMSCs,免疫组化技术检测UCMSCs表面特异标志物表达;四组细胞分别加入单纯培养液空白对照组（A组）、单纯生长因子诱导组（B组）、单纯黄芪诱导组（C组）和黄芪联合生长因子诱导液（D组）,倒置显微镜下观察其形态并检测NSE、NF和GFAP的阳性表达,结果进行统计学分析.结果 人UCMSCs表面标记CD29、CD44、CD105强阳性,CD106弱阳性,CD34、CD45阴性,符合人UCMSCs的特征;人UCMSCs经黄芪诱导后细胞表面GFAP、NSE均为强阳性表达.结论 人UCMSCs可以在体外采用组织块贴壁培养法培养成功,黄芪可将人UCMSCs成功的诱导为神经样细胞,为组织化人工神经的制备和周围神经缺损的治疗提供了新的方法.

Objective To investigate the biological characteristics of human umbilical cord derived mesenchymal stem cells（UCMSCs） and inducing them to differentiate into neurons in vitro,in order to provide stem cell resource for the tissue engineering artificial nerve. Methods UCMSCs were cultured from Wharton jelly of human umbilical cord in the condition of sterilitas,surface antigens of UCMSCs were detected by immunohistochemistry.The frist group of cells were added by virgin nutrient,the second group of cells were induced by merely growth factors,the third of cells were induced by merely astragalus mongholicus and the fourth by the astragalus mongholicus with growth factor,observed the morphology of the cells of the two groups under inverted microscope,and determine the positive expression rate of nerve cells＇ markers,such as NSE,NF and GFAP.The results were statistically analyzed.Results UCMSCs were strongly positive for CD29,CD44,CD105,weakly positive for CD105 and negative for CD34,CD45.After induced by Astragalus mongholicus,UCMSCs were strongly positive for GFAP,NSE. Conclusion UCMSCs can be successfully cultured from the adherent tissue pieces,Astragalus mongholicus can successfully induce them to differentiate into neurons in vitro,to provide a new method of making the tissue engineering artificial nerve and treat theperipheraly nervus defect.