Maintenance of the differentiated type II cell phenotype by culture with an apical air surface.
The study of differentiated functions of alveolar type II cells has been hampered because of the lack of good in vitro systems. We report that culture of type II cells on collagen gels with an apical surface exposed to air promotes expression of differentiated type II cell characteristics. Cells cultured in this manner are cuboidal, contain lamellar bodies, and produce tubular myelin; in addition, they secrete phosphatidylcholine in response to exogenous ATP. Cultures contain mRNA for surfactant proteins A, B, and C and surfactant proteins A, B, and D. In contrast, when type II cells are cultured with an apical surface exposed to liquid rather than to air, the cells are squamous, do not express surfactant proteins or their respective mRNA, and do not contain lamellar bodies or produce tubular myelin. Type II cells cultured on plastic for 7 days, which no longer express mRNA for surfactant proteins, can be induced to express these mRNA by changing culture conditions to that of an air surface. The culture system described in this paper should be useful for studies of surfactant metabolism, regulation of alveolar epithelial phenotypic expression, and the processing of transiently expressed transgenes.
Dobbs
L G
LG
Department of Medicine, University of California San Francisco 94143, USA.
Pian
M S
MS
Maglio
M
M
Dumars
S
S
Allen
L
L
D000388:Air; D000818:Animals; D015415:Biomarkers; D002454:Cell Differentiation; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry; D003598:Cytoskeletal Proteins / Q000037:antagonists & inhibitors; D005856:Germ-Free Life; D008168:Lung / Q000166:cytology* / Q000378:metabolism / Q000502:physiology*; D010641:Phenotype; D010713:Phosphatidylcholines / Q000378:metabolism; D010969:Plastics; D011663:Pulmonary Surfactants / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D013499:Surface Properties
D015415:Biomarkers; D003470:Culture Media; D003598:Cytoskeletal Proteins; D010713:Phosphatidylcholines; D010969:Plastics; D011663:Pulmonary Surfactants; D012333:RNA, Messenger