Bronchial epithelial spheroids: an alternative culture model to investigate epithelium inflammation-mediated COPD.
BACKGROUND
Chronic obstructive pulmonary disease (COPD) is characterized by abnormal lung inflammation that exceeds the protective response. Various culture models using epithelial cell lines or primary cells have been used to investigate the contribution of bronchial epithelium in the exaggerated inflammation of COPD. However, these models do not mimic in vivo situations for several reasons (e.g, transformed epithelial cells, protease-mediated dissociation of primary cells, etc.). To circumvent these concerns, we developed a new epithelial cell culture model.


METHODS
Using non transformed non dissociated bronchial epithelium obtained by bronchial brushings from COPD and non-COPD smokers, we developed a 3-dimensional culture model, bronchial epithelial spheroids (BES). BES were analyzed by videomicroscopy, light microscopy, immunofluorescence, and transmission electron microscopy. We also compared the inflammatory responses of COPD and non-COPD BES. In our study, we chose to stimulate BES with lipopolycaccharide (LPS) and measured the release of the pro-inflammatory mediators interleukin-8 (IL-8) and leukotriene B4 (LTB4) and the anti-inflammatory mediator prostaglandin E2 (PGE2).


RESULTS
BES obtained from both COPD and non-COPD patients were characterized by a polarized bronchial epithelium with tight junctions and ciliary beating, composed of basal cells, secretory cells and ciliated cells. The ciliary beat frequency of ciliated cells was not significantly different between the two groups. Of interest, BES retained their characteristic features in culture up to 8 days. BES released the inflammatory mediators IL-8, PGE2 and LTB4 constitutively and following exposure to LPS. Interestingly, LPS induced a higher release of IL-8, but not PGE2 and LTB4 in COPD BES (p < 0.001) which correlated with lung function changes.


CONCLUSIONS
This study provides for the first time a compelling evidence that the BES model provides an unaltered bronchial surface epithelium. More importantly, BES represent an attractive culture model to investigate the mechanisms of injuring agents that mediate epithelial cell inflammation and its contribution to COPD pathogenesis.
Service des Maladies Respiratoires, Hôpital Maison Blanche, CHU de REIMS, France. gdeslee@chu-reims.fr
Gaetan
G
Deslee
Sandra
S
Dury
Jeanne M
JM
Perotin
Denise
D
Al Alam
Fabien
F
Vitry
Rachel
R
Boxio
Sophie C
SC
Gangloff
Moncef
M
Guenounou
François
F
Lebargy
Abderrazzaq
A
Belaaouaj
D001980:Bronchi / Q000473:pathology*; D018929:Cell Culture Techniques / Q000379:methods; D006801:Humans; D011014:Pneumonia / Q000473:pathology; D029424:Pulmonary Disease, Chronic Obstructive / Q000473:pathology*; D020545:Respiratory Mucosa / Q000473:pathology*; D018874:Spheroids, Cellular