Metabolic properties and ultrastructure of alveolar type II cells isolated with elastase.
We used porcine pancreatic elastase to isolate type II cells from the lungs of rats; the yield and purity of the type II cells was better than that obtained by methods using trypsin. In 102 experiments we obtained 82 +/- 23 . 10(6) cells/rat, 68 +/- 11% (mean +/- S.D.) of which type II cells. This preparation of cells, when centrifuged over a discontinuous density gradient, yielded 25 +/- 10 . 10(6) cells/rat, 80 +/- 13% of which were type II cells (n = 102). The cells, after density gradient centrifugation, could be futher purified by centrifugal elutriation (94 +/- 3% type II cells, n = 22) or adherence in primary culture (94 +/- 2% type II cells, n = 34). Type II cells isolated with elastase are similar morphologically and biochemically to type II cells isolated  from rats  with trypsin. The preparations of cells appeared healthy by several different criteria: ultrastructure, exclusion of vital dye, lack of stimulation of oxygen consumption by exogenous sodium succinate, and linear rates of oxidation of [1-14C]palmitic acid and of incorporation of [1-14C]acetate into fatty acids. Type II cells consumed 75 +/- 20 nmol O2/10(6) cell per h, oxidized [1-14C]palmitic acid at a rate of 0.4 nmol/10(6) cells per h, and incorporated [1-14C]acetate into fatty acids at a rate of 7.5 nmol/10(6) cells per h.
D000085:Acetates / Q000378:metabolism; D000818:Animals; D002469:Cell Separation; D005227:Fatty Acids / Q000378:metabolism; D008297:Male; D008854:Microscopy, Electron; D010101:Oxygen Consumption; D010169:Palmitic Acids / Q000378:metabolism; D010196:Pancreatic Elastase; D011650:Pulmonary Alveoli / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats
D000085:Acetates; D005227:Fatty Acids; D010169:Palmitic Acids; D010196:Pancreatic Elastase