摘要
目的:研究不同剂量的3H-TdR对骨髓间充质干细胞生长代谢的影响,以确定安全的3H标记剂量。方法:在骨髓间充质干细胞培养瓶加入不同浓度的3H-TdR,描绘各细胞生长曲线,用Hochest染色观察细胞凋亡情况及用流式细胞仪检测细胞周期和骨髓间充质干细胞特有细胞表面标志物CD29、CD166、CD44、CD45的变化,观察对干细胞SOD、MDA的影响。结果:研究发现以4μCi/2ml浓度的3H-TdR标记干细胞时干细胞生长受到抑制,3μCi/2ml以上的浓度细胞均有不同程度的凋亡,流式细胞仪发现高浓度的核素能使干细胞被阻滞在S期及G2-M期,而且其细胞内的SOD下降及MDA升高,上述变化都有随核素剂量变化而变化的趋势;而用1μCi/2ml和正常组差异无统计学意义(P>0.05),标记过的骨髓间充质干细胞连续传代培养其增殖能力不受影响。结论:较高浓度的3H-TdR对干细胞的培养、示踪有影响,而1μCi/2ml的3H-TdR进行示踪研究是安全的。
Objective：To study the cellular toxicity of 3H-TdR in labeling mesenchymal stem cells（MSCs）.Methods： Different dosage of 3H-TdR was added to the culture bottles.The growth curves of BMSC in different concentrations were made,the apoptosis of cell was observed by Hochest stain and the cell cycle was observed with flow cytometer.The cell surface marks of MSCs（CD29,CD166,CD44,CD45） were detected by flow cytometer and the influence on SOD or MDA level was observed.Results： The researches showed that MSCs growth was suppressed at 4μCi/2ml concentration of 3H-TdR.High concentration of 3H-TdR caused MSCs to be arrested at the S and G2-M stage;MSCs showed obvious apoptosis when the concentration of 3H-TdR exceeded 3μCi/2ml;SOD decreased,MDA increased and their changes were in proportion to the alteration of concentration of 3H-TdR.The proliferation ability of successive progeniture was not affected by 3H-TdR when its concentration didn＇t exceed 1μCi/2ml.Conclusion： Higher concentration of 3H-TdR can arrest cell cycle of stem cell,affect its growth and tracer function,but concentration lower than 1μCi/2ml has no effect on biological characteristic of stem cell.Therefore it is safe for tracing study.
出处
《泸州医学院学报》
2013年第3期211-215,共5页
Journal of Luzhou Medical College
基金
国家自然基金资助项目(30370510)
