The preliminary exploration of Calu-3 cell air-liquid interface culture and exposure research / 中国药理学通报

ABSTRACT

Aim To explore the air-liquid interface(ALI)culture conditions of Calu-3 cells and their tolerance to exposure to clean air in the exposure system. Methods Calu-3 cells were cultured in three stagesflask expansion,liquid-liquid culture until full Transwell membrane covered,and ALI culture,and the cell barrier status was determined by cell trans-epithelium electrical resistant(TEER)measurement and expression of tight junction protein ZO-1; Calu-3 cells were exposed to clean air using the air-liquid interface exposure system for 1,2,3,4,5,6 hours,and cells cultured in incubators were used as controls to detect changes in Calu-3 cell activity and TEER after exposure,and to determine the single maximum exposure time of Calu-3 cells in the in vitro exposure system based on ALI culture. Results Calu-3 cells were cultured in liquid-liquid culture for 8±1 days to grow full Transwell membranes and barrier formation,and then were transferred to ALI culture. The barrier was intact and in a stable state for 1 to 18 days of ALI culture,and could be used for exposure experiments. The activity of Calu-3 cells decreased significantly after 6 hours of clean air exposure(P<0.01),and TEER decreased significantly after 4,5,and 6 hours of clean air exposure(P<0.05,P<0.01,P<0.01). Conclusions Calu-3 cells cultured for 1 to 18 days in ALI can be used for air-liquid interface exposure experiments; the combined changes in cell activity and TEER suggest that the maximum single exposure time for Calu-3 cells exposed to clean air in the exposure system is 3 hours.