Am. J. Respir. Cell Mol. Biol., Vol 15, No. 3, 09 1996, 312-318.
Morphologic and functional properties of bronchial cells isolated from normal and asthmatic subjects
F Goulet, LP Boulet, J Chakir, N Tremblay, J Dube, M Laviolette, M Boutet, W Xu, L Germain and FA Auger
Laboratoire d'Organogenese Experimentale, Hopital du Saint-Sacrement, Quebec City, Quebec, Canada.
Recent advances in biomedical sciences have led to the development of
various methods for the evaluation of the physiopathology of respiratory
diseases. This study reports morphologic and functional features of cells
isolated by a new method from bronchial biopsies of normal and asthmatic
subjects. Both epithelial and fibroblastic cells were isolated from the
same biopsies using collagenase. The cells were cultured for several
passages and stored frozen. Two selective culture media were used in order
to obtain pure epithelial and fibroblastic cell populations.
Immunofluorescence analysis of intermediate filaments, keratins, and
vimentin confirmed the type of the isolated cells. The proportions of
alpha-actin-expressing cells varied among the fibroblastic cell populations
isolated from normal and asthmatic subjects. Interestingly, the population
containing high numbers of alpha-actin-expressing cells and presenting the
fastest collagen contraction kinetic was isolated from bronchial biopsies
of an asthmatic subject. Moreover, the fibroblastic cells that showed the
best contractile properties 24 h after their seeding in floating collagen
gels were isolated from bronchial biopsies of asthmatic patients having
PC20 values below 1 mg/ml. On the basis of these data, we propose a new
approach to isolate, culture and characterize human bronchial cells in
vitro.
