The effect of substratum and serum on the lipid synthesis and morphology of alveolar type II cells in vitro.
To determine the effect of various culture conditions on the maintenance of lipid synthesis and morphology in alveolar type II cells, we cultured isolated adult rat alveolar type II cells on either plastic or denuded human amnionic basement membrane (ABM) in medium supplemented with either fetal bovine, porcine, horse, rat, or human serum. Lipid synthesis was assessed by incubation with [1-14C]acetate and determination of the distribution of radiolabel into individual lipid classes. Cells cultured on ABM incorporated significantly higher percentages of acetate into either phosphatidylcholine (PC) or phosphatidylglycerol (PG), and retained lamellar inclusions and a more characteristic cuboidal shape for longer periods than did cells cultured on plastic. Compared to other sera, cells cultured in the presence of rat serum incorporated the highest percentages of acetate into PC and saturated PC, had the best preservation of lamellar-body ultrastructure, and also appeared to contain more multivesicular bodies. The percent composition of linoleic acid, an essential fatty acid, was found to vary widely among the different sera. Supplementing media with linoleic acid resulted in a marked increase in acetate incorporation into saturated PC and a decreased incorporation into PG. We conclude that for maintenance of differentiated function of adult rat alveolar type II cells in primary culture (1) ABM is preferable to plastic as a culture substratum, (2) rat serum is preferable to fetal bovine serum as a serum supplement, and (3) the regulation of lipid synthesis by linoleic acid causes disparate effects on PG and saturated PC synthesis.
Cott
G R
GR
Department of Medicine, National Jewish Center for Immunology and Respiratory Medicine, Denver, Colorado 80206.
Walker
S R
SR
Mason
R J
RJ
D000818:Animals; D001485:Basement Membrane; D001769:Blood; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology; D003584:Cytological Techniques; D008055:Lipids / Q000032:analysis / Q000096:biosynthesis*; D011650:Pulmonary Alveoli / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D011663:Pulmonary Surfactants / Q000096:biosynthesis; D051381:Rats; D011919:Rats, Inbred Strains; D013045:Species Specificity
D003470:Culture Media; D008055:Lipids; D011663:Pulmonary Surfactants