关键词: 关键词：成年小鼠, 肺成纤维细胞, 分离, 纯化, 原代细胞培养

Abstract: Abstract: Objective To explore a reasonable method for isolation, purification and primary culture of lung fibroblasts from adult mouse lungs and research the growth curve of lung fibroblast. Methods Adult mice lung fibroblasts were isolated and cultured using three methods of lung tissue directly adherent, trypsin digestion, mixed culture method of lung tissue adherent and trypsin digestion. Lung fibroblasts were purified in different centrifugal force and different adherence. Lung fibroblasts were identified according to cell shape and the expression of vimentin and α-SMA by immunohistochemistry and optical microscope. Their growth curve was measured by MTT. Results Single purified lung fibroblasts were elongated spindle-shaped, cord-like. Monolayer covered lung fibroblasts were swirling, palisading, radially arranged. The expression of vimentin in lung fibroblasts was positive and that of α-SMA was weakly positive. Cells were appeared exponential growth after seeded 48 h and reached their peak at 120 h. Conclusions Lung fibroblasts from adult mouse were successful isolated, purified and cultured by above three methods. This study provided reliable and feasible technical approaches to obtain adult mouse lung fibroblasts.

Key words: Key words: Adult mouse, Lung fibroblasts, Isolation, Purification, Primary cell culture