关键词: 树鼩, 肺成纤维细胞, 鉴定, 传代培养

Abstract: Objective To establish a reliable method for isolation, identification and subculture of pulmonary fibroblasts in tree shrew. Methods Pulmonary fibroblasts of neonatal tree shrew were isolated and cultured by tissue adherence culture and trypsin digestion respectively. The fibroblasts was observed by inverted microscope and identified by immunofluorescence with Vimentin. Then the pulmonary fibroblasts was subcultured, and cryopreservation and recovery experiments were carried out. The cell growth curve was drawn by MTS method, and human embryonic lung diploid fibroblasts (KMB-17) were used as control. Results Tree shrew pulmonary fibroblasts could be obtained by both tissue adherence culture and trypsin digestion. The cells were mainly spindle-shaped, and vimentin immunofluorescence assay were generally positive, which were consistent with KMB-17. Tree shrew pulmonary fibroblasts can be subcultured 4-5 times continuously. After cryopreservation and recovery, the cells can still remain viable and enter the logarithmic growth phase about 3 days later with a cell density of 5×10⁴ cells/mL. Conclusion The pulmonary fibroblasts in tree shrew can be effectively obtained by tissue adherence and trypsin digestion, which will be very helpful for the study of lung-related diseases in vitro.

Key words: Standardization and Application Research in Tree Shrew, Kunming 6500118, China)