Abstract
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 non-specific 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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Adamson IYR, Bowden DH, Cote MG, Witschi H (1977). Lung injury induced by butylated hydroxytoluene. Lab Invest 36: 26–32.
Adamson IY, Prieditis HL (1995). Response of mouse lung to carbon deposition during injury and repair. Environ Hlth Perspect 103: 72–76.
Baron J, Voigt JM (1994). Localization, distribution and induction of xenobiotic metabolising enzymes and aryl hydrocarbon hydroxylase activity within the lung. In: Gram TE (ed) Metabolic activation and toxicity of chemical agents to lung tissue and cells, pp 41–76. Oxford: Pergamon Press.
Bingle L, Bull TB, Fox, Guz A, Richards RJ, Tetley TD (1990). Type II pneumocytes in mixed cell culture of human lung; A light and electron microscopic study. Environ Hlth Perspect 85: 71–80.
Devereux TR, Domin BA, Philpot RM (1994). Xenobiotic metabolism by isolated pulmonary bronchiolar and alveolar cells. In: Gram TE (ed), Metabolic activiation and toxicity of chemical agents to lung tissue and cells, pp 25–40. Oxford: Pergamon Press.
Devereux TR, Fouts JR (1980). Isolation and identification of Clara cells from rabbit lung. In vitro. 16: 958–968.
Edelson JD, Shannon JM, Mason RJ (1989). Effects of two extracellular matrices on morphologic and biochemical properties of human type II cells in vitro. Am Rev Respir Disease 140: 1398–1404.
Evans WA, Hext PM, Richards RJ (1995). Isolation and short-term culture of rat olfactory cells enriched with NADPH-dependent reductase activity. In Vitro Toxicol 8(1): 87–95.
Haagsman HP (1994). Surfacant proteins A and D. Biochemical Society Transactions 22: 100–106.
Harwood JL, Desai R, Hext R, Tetley T, Richards RJ (1975). Characterisation of pulmonary surfactant from ox, rabbit, rat and sheep. Biochem J 151: 707–714.
Hoet PHM, Lewis CPL, Demetz M, Nemery B (1994). The uptake of putrescine in the presence of paraquat in human lung slices and isolated alveolar type II cells. Biochem Pharmacol 48: 517–524.
Jones HE, Blundell GK, Wyatt I, John RA, Farr SJ and Richards RJ (1992). The accumulation of pentamidine into rat lung slices and its interaction with putrescine. Biochemical Pharmacology 43: 431–437.
Kikkawa Y, Yoneda K (1974). The type II epithelial cell of the lung. I. Method of isolation. Lab Invest 30: 76–84.
Lessire F, Gustin P, Delaunois A, Vloden S, Nemmar A, Vargas M, Ansay M (1996). Relationship between parathione and paraoxon toxicokinetics, lung metabolic activity, and cholinesterase inhibition in guinea pig and rabbit lungs. Toxicology and Applied Pharmacology 138: 201–210.
Lesur O, Arsalane K, Lane D (1996). Lung alveolar epithelial cell migration in vitro: modulators and regulation processes. Am J Physiol 270: L311-L319.
Lewis RW, Richards RJ, Harwood JL (1987). Pulmonary surfactant biosynthesis: studies in vivo and in vitro. Biochemical Society Transactions 15: 480–481.
McGowan SE (1992). Extracellular matrix and the regulation of lung development and repair. FASEB J 6: 2895–2904.
Miller BE, Hook GER (1990). Hypertrophy and hyperplasia of alveolar type II cells in response to silica and other pulmonary toxicants. Environ Hlth Perspect 85: 15–23.
Nemery B, Hoet PHM (1993). The use of isolated lung cells in pulmonary toxicology. Toxicol in Vitro 7: 359–364.
Oreffo VIC, Morgan A, Richards RJ (1990). Isolation of Clara cells from the mouse lung. Environ Health Perspect 85: 51–64.
Papadopoulos T, Inoescu L, Dammrich J, Toomes H, Muller-Hermelink HK (1990). Type I and Type IV collagen promote adherence and spreading of human type II pneumocytes in vitro. Lab Invest 62: 562–569.
Richards RJ, Davies N, Atkins J, Orefoo VIC (1987). Isolation, biochemical characterisation, and culture of lung type II cells of the rat. Lung 165: 143–158.
Robinson PC, Voelker DR, Mason RJ (1984). Isolation and culture of alveolar type II epithelial cells. Characterisation of their phosphalipid secretion. Am Rev Respir Disease 130: 1156–1160.
Smith LL, Nemery B (1992). Cellular specific toxicity in the lung. In: De Matteis F, Lock EA (eds), Selectivity and molecular mechanisms of toxicity, pp 3–26. London: MacMillan Press.
Voorhourt WF, Weaver TE, Haagsman HP, Geuze HJ, Vangolde LMG (1993). Biosynthetic routing of pulmonary surfactant proteins in alveolar type II cells. Microsc Res Tech 26: 366–373.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Murphy, S.A., Dindsale, D., Hoet, P. et al. A comparative study of the isolation of type II epithelial cells from rat, hamster, pig and human lung tissue. Methods Cell Sci 21, 31–38 (1999). https://doi.org/10.1023/A:1009825008023
Issue Date:
DOI: https://doi.org/10.1023/A:1009825008023