400mL·L^(-1)氧气对体外培养的人胚肺成纤维细胞增殖的促进作用

Effects of 400 mL·L^(-1) Oxygen on Proliferation of Human Embryo Lung Fibroblasts Cultured in Vitro

目的探讨400 mL·L-1O2对体外培养的人胚肺成纤维细胞(HELFs)增殖的促进作用。方法将生长良好的HELFs按13传代后,用无血清培养基培养24 h,分别置于空气及400 mL·L-1O2的细胞培养箱中培养5 d,每天收集细胞并计数、绘制细胞生长曲线;于培养后1 d、3 d及5 d,作Giemsa液染色,计算细胞分裂指数;培养后5 d收集细胞,流式细胞术检测细胞周期。结果细胞生长曲线显示400 mL·L-1O2较空气利于细胞生长;400 mL·L-1O2干预细胞3 d、5 d后细胞分裂指数分别为2.47%和2.03%,均明显高于空气组(1.90%和1.65%)(P<0.01,0.05);与空气组比较,400 mL·L-1O2干预细胞5 d后,HELFs处于G0/G1期细胞的比率明显减少(P<0.01),S期细胞比率显著增加(P<0.01),G2/M期细胞比率无明显差异(P>0.05)。结论 400 mL·L-1O2促进体外培养的HELFs的生长和增殖。

Objective To explore the direct effect of 400 mL ·L-1 oxygen （02 ） on proliferation of human embryo lung fibroblasts （HELFs） cultured in vitro. Methods After HELFs were passaged according to the ratio of 1 to 3 and cultured 24 hours in DMEM with no fe- tal bovine serum,then randomly divided into air group and 400 mL·L-1O2 group. Cells were cultured for5 days. Cells were collected every- day and cellular growth curve was drew;cell division was determined by Giemsa staining after cultured 1 day,3 days and 5 days;the changes of cell cycle were analyzed by flow cytometry after cultured 5 days. Results The concentrations of 400 mL ·L-1O2 promoted cell growth. As compared with air group, the division index was 2.47% and 2.03% in 400 mL ·L-1O2 group, 1.90% and 1.65% J n air group after treated cells 3 d and 5 d （ P 〈 0.01,0.05 ）. As compared with air group, the fraction of cells in G0/G1 phase decreased （ P 〈 0.01 ） and that in S phase increased （ P 〈 0.01 ） in 400 mL ·L-1O2 group, the fraction of cells in G2/M phase had no significant difference （ P 〉 0.05 ）. Conclusion The concentration of 400 mL·L-1 promote HELFs growth and proliferation.