低氧对人脐带间充质干细胞生物学特性及增殖的影响  

Effect of hypoxia on biological characteristics and proliferation of human umbilical cord mesenchymal stem cells

作　　者：宋希拿 孟恒星 张宇光[1,2] 武文杰[1,2] 李茜[1,2] 韩俊领[1,2] 

机构地区：[1]中国医学科学院.北京协和医学院血液病医院.血液学研究所,天津300020 [2]协和干细胞基因工程有限公司天津市脐带血造血干细胞库,天津300384

出　　处：《生物医学工程与临床》2014年第2期106-112,共7页Biomedical Engineering and Clinical Medicine

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.

关 键 词：人脐带间充质干细胞 低氧 细胞增殖 细胞凋亡 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学]