小鼠脂肪间充质干细胞体外培养时间与细胞衰老关系初探  

The relationship between cell culture time and cell senescence of mouse adiposederived mesenchymal stem cells in vitro

作　　者：李琼[1] 左红波[1] 张珉[2] 李慈霞[1] 郭志坤[1] 

机构地区：[1]新乡医学院河南省医用组织再生重点实验室,新乡453003 [2]河南省肿瘤医院肝胆外科,郑州450000

出　　处：《中国免疫学杂志》2013年第10期1028-1032,共5页Chinese Journal of Immunology

基　　金：河南省杰出创新人才计划(084200410020);教育厅基础研究项目(12A310003)资助

摘　　要：目的:探讨脂肪间充质干细胞(Adipose-derived mesenchymal stem cells,ADMSCs)体外培养时间与细胞衰老之间的关系,为细胞治疗提供实验参考。方法:分离、培养小鼠ADMSCs,MTT法绘制细胞生长曲线;β-半乳糖苷酶(SA-β-Gal)染色检测第1至第10代ADMSCs以及第4代细胞培养第1至第8天细胞的衰老情况;流式细胞术检测细胞周期变化;电镜观测第3和第8代细胞的超微结构。结果:衰老ADMSCs胞浆呈蓝色,随着代次增加细胞衰老率升高,第8代开始高于第1代细胞(P<0.05),并且G0/G1期细胞所占比例升高,S期和G2/M期细胞所占比例明显降低,提示细胞分裂增殖速度减慢;相同代次细胞,随着培养天数增加细胞衰老率呈上升趋势,第4天开始细胞的衰老比率明显高于第1天(P<0.05)。随着细胞代次增加,细胞膜出现较多突起,且胞质内脂滴增多,出现自噬体。结论:ADMSCs细胞衰老率与体外培养时间正相关。Objective：Investigation of the relationship between culture time and cell senescence of adipose-derived mesenchy- real stem cells （ADMSCs） in vitro to provide experimental evidence for cell therapy. Methods： Isolated and cultured mouse ADMSCs and growth curve of ADMSCs was made by MTT method. Senescence index of ADMSCs from 1th passage to 10th passage were measured by SA-β-Gal staining. The cell cycles were detected by flow cytometry. Ultrastructure of ADMSCs was observed by transmission elec- tron microscope. Results： The cytoplasm of aging ADMSCs was expressed blue, and index of cell senescence was positive correlation with culturing time in vitro. With increasing of passages, cell senescence index of the 8~h generation was higher than that The first generation cells （P 〈0.05）. The percentage of ADMSCs in G0/G1 was increased with increasing of culturing time, in contrast, the percentage of ADMSCs in G2/M was deereased,that means the speed of division growth was descend. With increasing of the culture days, the index of cell senescence was upward trend, and the index of cell senescence on 4th day was significantly higher than that in the first day （ P 〈 0.05 ）. With increasing of passages, there were more big scabrosities and many microspikes on cell membrane and many lipid droplets and autophagosomes in cytoplasm. Conclusion： Index of cell senescence about ADMSCs was positive correlation with culturing time in vitro.

关 键 词：脂肪间充质干细胞 细胞衰老 超微结构 

分 类 号：R392[医药卫生—免疫学]