关键词: 鼻息肉, 原代培养, 低氧, 成纤维细胞

Abstract: Objective    To investigate the primary culture and identification of fibroblasts cells from nasal polyps under normoxia and hypoxia for further study on their biological features. Methods    The specimens of nasal polyps(20 cases)endoscopically removed in our department from 2012 to 2013 were placed into 4℃ sterile phosphate-buffered saline(PBS)solution supplemented with penicillin and streptomycin, repeatedly washed, immersed, cut into pieces, and tubed into two portions to EP, respectively with 0.1％collagenase Ⅰ and 1mL of dispase Ⅱ overnight. The next day, the samples were removed, pipetted slightly to detach the epithelia, after to count, and added in broth (DMEM+10% fetal bovine serum+1×105U/L penicillin+100mg/L streptomycin), respectively in normoxia and hypoxia conditions. Besides normoxia training with air as the mixed gas, oxygen volume fraction of about 20%; hypoxia training with nitrogen as the mixed gas, regulating oxygen volume fraction of 5%, the other culture conditions were the same with the volume fraction of 5% CO2 and saturated humidity at 37℃. Under an inverted phase microscope, the cells morphology, proliferation, growth and infection were observed. The proliferation of cells was tested by CCK-8. Results    Overall improvement was observed in the number of cells in dispaseⅡ and was higher than that of type collagenase Ⅰ(P<0.05). Nasal polyp fibroblasts in normoxia and hypoxia condition training had no obvious difference. Conclusion    DispaseⅡ can be used to obtain more cells. In normoxia and hypoxia condition, the above mentioned model is more suitable for primary culture due to its fast proliferation, stable and reliable cell supply for nasal polyps research.

Key words: Fibroblasts, Primary culture, Hypoxia, Nasal polyps