低氧培养增强骨髓源内皮祖细胞的增殖、黏附、迁移和生存能力

Hypoxia condition can enhance proliferation, adhesion, migration, and viability abilities of bone morrow-derived endothelial progenitor cells

目的评估低氧(1%和5%氧浓度)环境对骨髓源内皮祖细胞及其促血管新生相关的增殖、黏附、迁移和生存能力的影响。方法利用密度梯度离心技术分离SD大鼠骨髓单个核细胞,向内皮祖细胞进行诱导分化、扩增、培养和鉴定。然后在不同氧浓度(1%、5%和21%)的环境中继续培养,于第3天和第7天时检测不同氧浓度下培养细胞的增殖能力、黏附能力、迁移能力及生存能力。结果内皮祖细胞在不同氧浓度(1%、5%及21%)的培养箱中培养时,无论是培养至第3天或是第7天时,1%和5%氧浓度下培养细胞的增殖能力、黏附能力、迁移能力及生存能力均明显优于21%氧浓度培养的细胞(均P<0.05);除了第3天时5%氧浓度下培养细胞的增殖能力优于1%氧浓度下培养细胞(P<0.05)以及第3天的黏附能力及第7天时在1%和5%氧浓度下培养细胞的增殖能力比较差异无统计学意义(P>0.05)外,其余(黏附能力、迁移能力及生存能力)无论是培养至第3天或是第7天均是1%氧浓度下培养细胞优于5%氧浓度(均P<0.05)。结论不同氧浓度对骨髓源内皮祖细胞增殖、黏附、迁移以及生存(抗凋亡和死亡)能力产生不同的影响,与21%氧浓度相比,适当时间的相对低氧浓度(1%和5%)培养能够显著增强内皮祖细胞的增殖、黏附、迁移及生存能力。

Objective To evaluate effect ofhypoxia condition （1% or 5% oxygen concentration） on proliferation, adhesion, migration, or viability ability of bone morrow-derived endothelial progenitor cells （EPCs）. Methods The bone marrow mononuclear cells of SD rat were acquired with density gradient centrifugation method. They were cultured, induced, and differentiated to the EPCs. Then they were cultured respectively in three different oxygen concentrations （1%, 5%, or 21%）. On the 3rd day and the 7th day, the effects of the different oxygen concentrations （1%, 5%, or 21%） on the EPCs＇ neovascularization characteristics （including proliferation, adhesion, migration, and viability abilities） were evaluated. Results Whether cultured for the 3rd day or 7th day, the proliferation, adhesion, migration, and viability abilities of the cultured cells in the 1% and 5% oxygen concentrations were significantly better than those of the cultured cells in the 21% oxygen concentration （all P〈0.05）. Except for the proliferation ability of the cultured cells in the 5% oxygen concentration was significantly better than that of the cultured cells in the 1% oxygen concentration （P〈0.05） on the 3rd day, and the adhesion ability on the 3rd day and the proliferation ability on the 7th day had no significantly differences, the other abilities （adhesion, migration, and viability abilities） of the cultured cells in the 1% oxygen concentration were significantly better than those of the cultured cells in the 5% oxygen concentration （all P〈0.05）. Conclusion Different oxygen concentration has an effect on proliferation, adhesion, migration, or viability ability of bone morrow-derived EPCs, appropriate hypoxia condition （1% or 5% oxygen concentration) can enhance these abilities.