A comparative study of the isolation of type II epithelial cells from rat, hamster, pig and human lung tissue.
Data is reported on the reproducibility and purity of alveolar type II cell isolations from 4 species. Human and pig type II cells were isolated using a tissue slice method to remove blood and contaminating cells, whilst rat and hamster cells were isolated using the method of protease instillation. All cells were purified on Percoll gradients and by differential attachment. Cell type purity was assessed by phase contrast microscopy, electron microscopy (EM), percentage of cells alkaline phosphatase (AP) positive and percentage of cells staining strongly for NADPH dependent nitro blue tetrazolium reductase (NBT). These enzymes are considered as markers for type II and Clara cells respectively. The purity of all cell preparations was enhanced following 24 h culture on a biomatrix and whilst plating efficiency was similar for all species, the human tissue consistently yielded the highest purity of type II cells. All cells with lamellar bodies did not contain AP, and activity was variable between species. Further studies are needed to determine if NBT is equally nonspecific as a cell marker enzyme. In summary, sufficient type II cells of high purity can be isolated thus permitting interspecies comparative studies to investigate the effects of selective and non-specific pulmonary toxins, but more specific marker enzymes are required to identify Type II and Clara cells.
Murphy
S A
SA
School of Molecular and Medical Biosciences, Cardiff University, UK.
Dinsdale
D
D
Hoet
P
P
Nemery
B
B
Richards
R J
RJ
D000469:Alkaline Phosphatase / Q000032:analysis; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002452:Cell Count; D002469:Cell Separation / Q000379:methods / Q000592:standards*; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D006224:Cricetinae; D004847:Epithelial Cells / Q000166:cytology* / Q000648:ultrastructure; D006801:Humans; D002479:Inclusion Bodies / Q000648:ultrastructure; D008854:Microscopy, Electron; D008858:Microscopy, Phase-Contrast; D009251:NADPH-Ferrihemoprotein Reductase / Q000032:analysis; D011650:Pulmonary Alveoli / Q000166:cytology*; D051381:Rats; D015203:Reproducibility of Results; D013045:Species Specificity; D013552:Swine
D015415:Biomarkers; C021848:NADPH-tetrazolium reductase; D009251:NADPH-Ferrihemoprotein Reductase; D000469:Alkaline Phosphatase