不同标本处理方法降低COPD模型SD大鼠肺成纤维细胞原代培养过程中污染率的实验研究

Effects of different specimen processing methods in reducing contamination rate during primary culture of lung fibroblasts in Sprague-Dawley rats withchronic obstructive pulmonary disease:an experimental study

目的:通过对比不同方法处理COPD模型大鼠肺组织,寻找适当方法减少在COPD模型大鼠细胞培养中的污染机会,提高细胞培养成功率。方法:将同一COPD模型大鼠肺组织分成两部分,分别用不同方法对其预处理,用组织块培养法进行原代细胞培养,观察第24h、72h及后续培养情况。结果:经过刮除肺叶及气管内膜的组织原代细胞培养组污染率较直接组织培养的要低,肺叶培养污染率为100%。结论:(1)刮除肺叶及气管内膜的方法有助于减少COPD模型大鼠原代细胞培养组织法的污染;(2)COPD模型大鼠肺成纤维细胞原代培养的污染来源主要为肺部感染灶。

Objective： To investigate proper methods which can reduce the probability of contamination during cell culture and improve the success rate of cell culture in rats with chronic obstructive pulmonary disease（ COPD） through comparing different methods for processing lung tissues from COPD rats. Methods： Lung tissues from COPD rats were divided into two parts and were treated with different methods. Primary cell culture was performed using the tissue block culture method,and the condition of cell culture at 24 and 72 hours and follow- up culture were observed. Results： Primary cell culture oftissues collected by excision of the lung lobe and tracheal intima yielded a lower contamination rate compared with direct tissue culture,and the contamination rate of lung lobe culture was100%. Conclusion： Excision of the lung lobe and tracheal intima helps to reduce contamination in primary cell culture for COPD rats. The source of contamination in primary culture of lung fibroblasts from COPD rats mainly comes from pulmonary infection.