Culture and differentiation of mouse tracheal epithelial cells at air-liquid interface
10.3969/j.issn.1001-1978.2021.02.024
- Author:
Xiao-Li JI
1
;
Yue HU
1
;
Yun-Huan SHENG
1
;
Li-Ming TANG
1
Author Information
1. NMPA Key Lab for Quality Analysis of Chemical Drug Preparations, Dept of Pharmacology and Toxicology, Shanghai Institute for Food and Drug Control, Center for Drug Safety Evaluation
- Publication Type:Journal Article
- Keywords:
air-liquid interface;
airway epithelium;
ciliated cells;
mucous cells;
trachea
- From:
Chinese Pharmacological Bulletin
2021;37(2):282-288
- CountryChina
- Language:Chinese
-
Abstract:
Aim Air-liquid interface (ALI) cultures of mouse tracheal epithelial cells (MTEC) are a well-established model to study airway epithelial cells. MTEC provides a powerful ap¬proach for the evaluation of the inhalation toxicological in vitro. Methods C57BL/6 mouse tracheal-bronchial epithelial cells were obtained by digestion with protease in cold temperature o- vemight, and the digestion time was optimized to ensure the quantity and viability of the obtained cells. The cells were cul¬tured into collagen coated Transwell inserts. Proliferating phase and air-liquid interface culture were promoted with different cul¬ture media. The expression of tight junction protein and cell trans-epithelial electrical resistance(TEER) were used to evalu¬ate the formation of tight junction between cells and the analysis of cell polarity. The cilia structure was confirmed by electron mi¬ croscopy and immunofluorescence. Results Highly purified and viable primary airway epithelial cells could be harvested and subcultured by our methods, including morphology and immuno- cytochemistry staining confirmed the expression of MUC5AC, a- tubulin, p-tubulin-IV and ZO-1. The development of tight-junc¬tions and epithelium were similar with pseudostratified ciliated columnar epithelium morphology. Conclusions A comprehen¬sive protocol for ALI culture was established, reproducing the characteristic pseudostratified ciliated columnar epithelium mor¬phology and physiological functions in vitro. The MTEC protocol provides a stable and reliable method for the isolation, mucocili¬ary differentiation and reproducing.
