蜕皮甾酮在体外条件下对人骨髓间充质干细胞增殖的影响  

Effect of ecdysterone on the proliferation of human mesenchymal stem cells in vitro

作　　者：吴昌昊[1] 吴旭[1] 付小兵[2] 周云峰[1] 张元中[1] 张子良[1] 

机构地区：[1]南方医科大学南方医院胸心血管外科,广东广州510515 [2]解放军总医院第一附属医院全军烧伤研究所创伤修复重点实验室,北京100048

出　　处：《南方医科大学学报》2010年第5期1180-1182,1184,共4页Journal of Southern Medical University

摘　　要：目的分离培养并鉴定人骨髓间充质干细胞(hMSC),观察不同剂量蜕皮甾酮(EDS)对hMSC增殖的影响。方法密度梯度离心法获取hMSC,检测第2代细胞CD44、CD105、CD34及CD29的表达以鉴定其为hMSC。设不同浓度EDS的细胞培养体系(10、25、50和100μg/ml)与对照组(单纯hMSC培养基,无EDS)作对比,MTT比色法测定hMSC的增殖活力,并行流式细胞周期观察。结果 EDS各浓度组hMSC的MTT吸光度值与对照组比较均有明显差异(P<0.01);EDS浓度为25μg/ml时hMSCMTT吸光度值显著高于其余各组(P<0.01);在EDS浓度为10、50、100μg/ml时,组间吸光度值未见明显差异(P>0.05),但显著高于对照组(P<0.01)。流式细胞周期显示EDS浓度为25μg/ml时hMSC培养体中细胞S期细胞所占比例、增殖指数均明显高于对照组。结论 EDS在一定浓度范围内对体外培养条件下的hMSC具有生长促进作用,该作用在EDS浓度为25μg/ml时最为显著,但是EDS促进hMSC增殖的作用不随剂量的增加而增加。Objective To investigate the effect of ecdysterone （EDS） on the proliferation of human bone marrow mesenchymal stem cells （hMSCs） in vitro. Methods hMSCs were isolated from human bone marrow cell suspension by density gradient centrifugation. The expression of integrins CD44, CD105, CD34 and CD29 were examined by immunocytochemical method. EDS at 10, 25, 50 or 100 μg/ml were added in hMSC culture system, using the routine culture medium for hMSCs as control. The cell viability were analyzed by MTT assay and the cell cycle changes were examined by flow cytometry. Results The optical density （OD） differed significant between the EDS treatment groups and the control group （P0.01）, and 25 μg/ml EDS group showed the highest OD value （P0.01） without significant differences among 10, 50 and 100 μg/ml EDS groups （P0.05）. Flow cytometry showed that treatment of the cells with 25 μg/ml EDS significantly increased the cell percentages in S and G2M phases and the proliferation index （PI） of the cells as compared with the control group. Conclusion Within a given concentration range, EDS can promote the proliferation of hMSCs in vitro, and this effect can be the most obvious at the concentration of 25 μg/ml. The effect of EDS in promoting the proliferation of hMSCs does not positively correlate to EDS concentration administered.

关 键 词：蜕皮甾酮 人骨髓间充质干细胞 细胞增殖 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学] R961[医药卫生—基础医学]