Heterocellular cultures of pulmonary alveolar epithelial cells grown on laminin-5 supplemented matrix.
The pulmonary alveolar epithelium consists of alveolar type I (AT1) and alveolar type II (AT2) cells. Interactions between these two cell types are necessary for alveolar homeostasis and remodeling. These interactions have been difficult to study in vitro because current cell culture models of the alveolar epithelium do not provide a heterocellular population of AT1 and AT2 cells for an extended period of time in culture. In this study, a new method for obtaining heterocellular cultures of AT1- and AT2-like alveolar epithelial cells maintained for 7 d on a rat tail collagen-fibronectin matrix supplemented with laminin-5 is described. These cultures contain cells that appear by their morphology to be either AT1 cells (larger flattened cells without lamellar bodies) or AT2 cells (smaller cuboidal cells with lamellar bodies). AT1-like cells stain for the type I cell marker aquaporin-5, whereas AT2-like cells stain for the type II cell markers surfactant protein C or prosurfactant protein C. AT1/AT2 cell ratios, cell morphology, and cell phenotype-specific staining patterns seen in 7-d-old heterocellular cultures are similar to those seen in alveoli in situ. This culture system, in which a mixed population of phenotypically distinct alveolar epithelial cells are maintained, may facilitate in vitro studies that are more representative of AT1-AT2 cell interactions that occur in vivo.
Isakson
Brant E
BE
Department of Zoology and Physiology, University of Wyoming, 16th and Gibbon Streets, Laramie 82071-3166, USA.
Seedorf
Gregory J
GJ
Lubman
Richard L
RL
Boitano
Scott
S
D000818:Animals; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002455:Cell Division; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D003470:Culture Media; D004847:Epithelial Cells / Q000166:cytology; D005353:Fibronectins / Q000378:metabolism; D008297:Male; D011650:Pulmonary Alveoli / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D000097794:Kalinin
D015815:Cell Adhesion Molecules; D003094:Collagen; D003470:Culture Media; D005353:Fibronectins; D000097794:Kalinin