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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13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 investigate the effects of 400 mL · L -1 O 2 on the proliferation of human embryonic lung fibroblasts (HELFs) in vitro. METHODS: HELFs were passaged at 13 and cultured in serum-free medium for 24 h. Cells were cultured in air and 400 mL · L-1 O2 for 5 days. Cells were harvested daily and counted. Cells were harvested The growth curve was obtained. The cells were stained with Giemsa stain on day 1, day 3 and day 5 after culturing, and the cell division index was calculated. Cells were collected 5 d after culture, and the cell cycle was detected by flow cytometry. Results The cell growth curve showed that 400 mL · L -1 O 2 was more effective than air in cell growth. The cell division index was 2.47% and 2.03% after 400 mL · L -1 O 2 intervention for 3 d and 5 d, respectively, % And 1.65%, respectively) (P <0.01, 0.05). Compared with the air group, the percentage of HELFs in the G0 / G1 phase was significantly decreased (P <0.01) after treated with 400 mL·L- (P <0.01). There was no significant difference in G2 / M cell ratio (P> 0.05). Conclusion 400 mL · L -1 O2 promotes the growth and proliferation of HELFs cultured in vitro.