低氧对脂肪间充质干细胞向雪旺细胞分化的影响  

Effect of hypoxia on the differentiation of adipose-derived mesenchymal stem cells into Schwann cell

作　　者：陈鸥[1,2] 胡旭琪[2] 郑旭浩[2] 蒋良福[3] 

机构地区：[1]温州市中西医结合医院骨科 [2]温州医科大学附属第二医院脊柱外科 [3]温州医科大学附属第二医院手外科,浙江省医学重点学科325024

出　　处：《中华移植杂志（电子版）》2014年第2期39-42,共4页Chinese Journal of Transplantation(Electronic Edition)

基　　金：浙江省自然科学基金(Y2100253);温州市科技计划项目(Y20130203)

摘　　要：目的探讨低氧对脂肪间充质干细胞(ADMSCs)向雪旺细胞(SCs)分化能力的影响。方法分离培养SD大鼠ADMSCs并用流式细胞仪、茜素红染色、油红O染色鉴定。将成功分离的ADMSCs随机分为3组:常氧诱导组,在常氧条件下(5%CO2,21%O2,37℃)诱导;低氧处理+常氧诱导组,低氧处理(5%CO2,0.5%O2,37℃)后在常氧条件下诱导;低氧诱导组,低氧条件下(5%CO2,0.5%O2,37℃)诱导。观察各组细胞形态,MTT法检测细胞增殖情况,免疫荧光染色和Western blot检测SCs标志物GFAP和S-100的表达。结果细胞分离后,经流式细胞仪分析可见细胞表面CD44阳性、CD45阳性、CD90阳性,茜素红及油红O染色均为阳性。MTT法检测结果:低氧处理+常氧诱导组A值为0.861±0.039,高于常氧诱导组0.837±0.017,差异具有统计学意义(P<0.05);低氧诱导组A值为0.931±0.041,均高于常氧诱导组和低氧处理+常氧诱导组(P均<0.05)。免疫荧光染色发现常氧诱导组和低氧处理+常氧诱导组大量细胞GFAP和S-100表达阳性,低氧诱导组仅少量细胞S-100和GFAP表达阳性。Western blot检测发现常氧诱导组S-100蛋白表达最高,低氧处理+常氧诱导组GFAP蛋白表达最高,低氧诱导组S-100蛋白、GFAP蛋白表达均最低。结论低氧抑制ADMSCs向SCs的分化,低氧处理后的ADMSCs在常氧条件下仍可向SCs分化。Objective To investigate the effect of hypoxia on the differentiation ability of adipose-derived mesenchymal stem cells （ ADMSCs ） into Schwann ceils （ SCs ）. Methods The ADMSCs were isolated from rats and identified by flow cytometry, alizarin red staining and oil red O staining. ADMSCs were divided into three groups including normoxic induction group, hypoxia treatment ＋ normoxic induction group and hypoxia induction group. After induction, cell morphological change was analyzed by inverted microscope and the proliferation in each group was estimated by MTT assay. Expression of GFAP and S-100 was detected by immunofluorescence and Western blot. Results ADMSCs were isolated from rats successfully. ADMSCs had the strongest and weakest proliferation ability in hypoxia induction group and normoxic induction group respectively. The expression of GFAP and S-100 was obviously higher in ADMSCs of normoxic induction group and hypoxia treatment ＋ normoxia induction group, while lower in hypoxia induction group. Conclusion Hypoxia can inhibit the differentiation of ADMSCs into SCs, but the stem cell characteristics was kept. Hypoxia treatedADMSCs can differentiate into SCs under normoxic condition.

关 键 词：低氧 脂肪间充质干细胞 雪旺细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]