低氧和高糖促进体外培养的人骨髓间充质干细胞的增殖和迁移  

Hypoxia and hyperglycemia promote the proliferation and migration of human marrow mesenchymal stem cells in vitro

作　　者：燕洁静[1] 王海燕[1] 王雨生[1] 高凡[1] 李娜[1] 张鹏[1] 

机构地区：[1]第四军医大学西京医院眼科,全军眼科研究所,陕西西安710032

出　　处：《细胞与分子免疫学杂志》2015年第5期639-643,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81300770);国家重点基础研究发展计划资助(2011CB510200)

摘　　要：目的建立人骨髓间充质干细胞(h MSC)低氧高糖培养的细胞模型,研究其增殖、凋亡和移行等生物学特征。方法采用密度梯度离心法提取培养健康成人h MSC。取第3代细胞,根据培养氧浓度及葡萄糖浓度培养基类型分为4组:常氧常糖组(210 m L/L O2加5.56 mmol/L葡萄糖培养液)、常氧高糖组(210 m L/L O2加30 mmol/L葡萄糖培养液)、低氧常糖组(50 m L/L O2加5.56 mmol/L葡萄糖培养液)和低氧高糖组(50 m L/L O2加30 mmol/L葡萄糖培养液)。CCK-8法检测细胞的增殖能力、TranswellTM法检测细胞在24 h时的移行能力、流式细胞术检测细胞在24 h时的凋亡情况。结果与对照组相比,24 h和48 h时,常氧高糖组、低氧常糖组、低氧高糖组中的h MSC增殖能力增强,低氧高糖联合作用组增殖能力最强。在24 h时,常氧高糖组、低氧常糖组、低氧高糖组h MSC的移行能力较对照组增强。结论低氧及高糖促进h MSC增殖和移行,而对凋亡并无影响。Objective To investigate the cell proliferation, apoptosis and migration of human marrow mesenchymal stem cells （hMSCs） under hypoxia and hyperglycemia. Methods The hMSCs from healthy adults were isolated and harvested by density gradient centrifugation followed by adherent cultures. The cells was divided into four groups： control group （210 mL/L O2 with 5.56 mmol/L glucose）, high glucose and normoxia group （210 mL/L O2 with 30 mmol/L glucose）, normal glucose and hypoxia group （50 mL/L O2 with 5.56 mmol/L glucose） and high glucose and hypoxia group （50 mL/L O2 with 30 mmol/L glucose）. The cells at the third passage were cultured in different groups. Then, we used cell counting kit-8 （CCK-8） to detect the proliferation of hMSCs, TranswellTM assay to examine the migration of hMSCs, and flow cytometry to observe the apoptosis of hMSCs at 24 hours. Results Compared with the control group, the proliferation rate of hMSCs increased in the other groups at 24 and 48 hours, and the maximum proliferation effect was found in high glucose and hypoxia group. Compared with the control group, the migration ability of hMSCs in the other groups was enhanced dramatically at 24 hours, whereas there was no significant difference in the apoptosis rate among the four groups. Condusion Both high glucose and hypoxia could improve the migration and proliferation of hMSCs, but they have no effect on apoptosis.

关 键 词：骨髓间充质干细胞 低氧 高糖 脉络膜新生血管 

分 类 号：Q253[生物学—细胞生物学] Q255