黄芪多糖诱导大鼠骨髓间充质干细胞向神经样细胞分化

Astragalus polysaccharides induced rat bone marrow mesenchymal stem cells to neural cells differentiation

目的探讨大鼠骨髓间充质干细胞(BMSCs)在黄芪多糖的诱导下向神经样细胞分化。方法采用密度梯度离心法和贴壁筛选法成功培养BMSCs,并采用流式细胞仪检测CD44、CD45表达并对细胞进行鉴定。按照诱导方式的不同分为对照组、化学方法诱导组、黄芪多糖组。于诱导后第3、5、7天分别进行细胞形态学观察;并采用免疫细胞化学法和免疫组织荧光检测神经微管蛋白-βⅢ(Tuj1)和胶质纤维酸性蛋白(GFAP)的表达。结果形态学观察:黄芪多糖组诱导BMSCs 12 h后出现形态学变化,逐步形成典型的神经样细胞;化学诱导组在加入诱导剂后细胞有突起伸出,并随诱导时间变化而增多,3 d可见到一些细胞漂浮脱壁,随时间延长死亡细胞逐渐增多。对照组无典型的神经样细胞形态。免疫组化和免疫荧光检测:Tuj1阳性表达,逐步形成典型的神经样细胞;GFAP阴性对照组未发现阳性细胞。结论黄芪多糖成功诱导大鼠BMSCs向神经样细胞分化。

Objective To explore the rat bone marrow mesenchymal stem cells ( BMSCs) under the induction of astragalus polysac-charide to the sample for neural cell differentiation .Methods BMSCs were separated by density gradient centrifugation and adherence screening methods.By detection CD44, CD45,the cells were identified.According to the different methods of induction , the cells were dev-ided into control, chemically induced(5 mmol/L,β-mercap toethanol ), astragalus polysaccharides(20 mg/L)groups.After the induction of 3rd, 5th and 7th day, the morphology of cells were observed .The expressions of neuronal class Ⅲβ-tubulin( Tuj1) and Glial fibrillary acidic protein( GFAP) were measured by immunocytochemistry and Western blot .Results Astragalus polysaccharides induced BMSCs morphologi-cal changes after 12 h,gradually formed a typical sample nerve cells .The cells of chemical induced group had swelled out after joining induc-ers, and increasing gradually;at 3rd day,some cells were floated off the wall , extended the dead cells gradually .The typical sample neural cell morphology was not found in control group .Tuj1 expression was positive and formed a typical nerve cell .There was no positive cells in GFAP negative control group .Conclusions Astragalus polysaccharides successfully induce rat BMSCs to neural cell differentiation .