CLD鼠肺成纤维细胞的原代培养及生物学行为研究

Primary culture and biological behaviour study of lung fibroblasts from CLD rats

目的建立一套可靠的CLD鼠肺成纤维细胞(lung fibroblasts,LFs)分离、纯化和培养技术,并对其生物学行为进行研究,为高氧致肺间质纤维化体外研究提供平台。方法足月新生Wister大鼠生后即分别持续吸入85%氧或空气,于7d、14d、21d分别进行鼠肺LFs原代培养。绘制细胞生长曲线、计算细胞分裂指数、流式细胞术检测细胞周期。结果与空气组相比,高氧组LFs增殖速度增快,处于分裂期的细胞数增多,细胞周期中处于G1期细胞的比率逐渐减少,S期细胞比率增加,且呈时间依赖关系。结论本方法培养的LFs可用于CLD肺间质纤维化发病机制的体外研究;高氧可诱发LFs细胞周期紊乱,细胞异常增生。

[Objective] To develop a reliable method for isolation, purification and primary culture of lung fibroblasts （LFs） from CLD rats and to study their biological behaviour in vitro for further research. [Methods] Full-time newborn rats were continuously exposed to oxygen （85%） or room air after birth, primary culture of LFs were made on day 7, 14, 21. Cellular growth curve was drew, cellular division indexes were calculated, the changes of cell cycle of LFs were analyzed by flow cytometry. [Results] Hyperoxia caused an increased cell proliferation of LF and an increased cell riumber in division phase. The fraction of ceils in G1 phase was increased and in S phase was reduced. [Conclusion] This technique might be a reliable method for primary culture of LFs and could be used for the study on pathogenesis of pulmonary fibrosis of CLD ; Hyperoxia induced cell cycle regulation disordered and uncontrolled growth of LFs.