摘要
目的探讨低氧培养条件对人脐带间充质干细胞(hUC-MSCs)的生长和增殖的影响。方法采用人脐带酶消化法分离培养hUC-MSCs,分别将细胞置入体积分数20%、10%、3%O2条件下培养,通过细胞成脂、成骨分化诱导及流式细胞仪检测其表面标志物,鉴定hUC-MSCs;绘制细胞生长曲线,用流式细胞仪检测细胞周期及凋亡情况。结果低氧条件培养hUC-MSCs具有成脂、成骨等多项分化的潜能;流式细胞仪检测呈CD105、CD73、CD90、CD44、HLA-ABC高表达,CD106、CD29、CD45、CD34、HLA-DR低表达;体积分数3%O2培养组与正常体积分数20%O2培养组及体积分数10%O2培养组相比(各组倍增时间分别为17.2 h、20.8 h、18.9 h),细胞增殖速度加快(P<0.05);G0/G1期细胞减少(各组G0/G1期细胞数分别为77.11%±3.89%、83.92%±5.59%、80.19%±5.16%),S期细胞增多(S期细胞数分别为15.73%±2.56%、10.91%±1.86%、13.31%±2.31%),增殖指数升高(P<0.05);细胞凋亡率降低(流式细胞仪检测各组凋亡率分别为13.41%±1.39%、20.83%±1.81%、19.11%±2.44%)(P<0.05)。结论体积分数3%O2持续培养可促进hUCMSCs的增殖,减少细胞凋亡。
Objective To investigate the effects of hypoxia on the biological characteristics and proliferation of human umbilical cord mesenchymal stem cells（hUC-MSCs）. Methods hUC-MSCs were isolated by enzyme digestion and cultured with oxygen concentration at 20%, 10% and 3%, respectively. The expression of hUC-MSCs surface antigens was detected by flow cytometry, and the seventh passage hUC-MSCs were incubated in osteogenous and adipogenic medium. The growth curve under different culture conditions was drawn respectively. The cell cycle and the apoptosis were analyzed by flow cytometry. Results The hUC-MSCs cultured under condition of hypoxia can differentiate into adipocytes and osteoblasts. Flow cytometry showed that the expression of CD105, CD73, CD90, CD44, HLA-ABC were high, while the expression of CD106, CD29, CD45, CD34, HLA-DR were low. The growth and proliferation of hUC-MSCs cultured under 3 %O2 hypoxia condition were significantly increased （doubling time of 17.2-hour） as compared with that under 20 % oxygen and 10 % oxygen conditions（doubling time of 20.8-hour and 18.9-hour, respectively）（P 〈 0.05）. Less cells were arrested at G0/G1 phase under 3 % O2 as compared with that under 20 % oxygen and 10 % oxygen conditions（77.11% ± 3.89 %, 83.92 % ± 5.59 % and 80.19 % ± 5.16 %, respectively）, while the cell numbers at S phase cells increased under 3 % 02 hypoxia in comparison with that under 20 % oxygen and 10 % oxygen conditions（15.73 % ± 2.56 %, 10.91% ± 1.86 % and 13.31% ± 2.31%, respectively）. The proliferation index increased （P 〈 0.05） and apoptotic rate decreased under 3 % O2 hypoxia when compared with that under 20 % oxygen and 10 % oxygen conditions（apoptotic rates being 13.41% ± 1.39 %, 20.83 % ± 1.81% and 19.11% ± 2.44 %, respectively）（P〈 0.05）. Conclusion It is demonstrated that culture at 3 % O2 concentration could enhance cell proliferation and reduce apoptosis of hUC-MSCs.
出处
《生物医学工程与临床》
CAS
2014年第2期106-112,共7页
Biomedical Engineering and Clinical Medicine
关键词
人脐带间充质干细胞
低氧
细胞增殖
细胞凋亡
human umbilical cord mesenchymal stem cells
hypoxia
cell proliferation
cell apoptosis
