Abstract
We have developed an improved isolation procedure for rat lung alveolar Type II (ATII) cells which yields a more representative sample of ATII cells with respect to their densities. This procedure includes an initial selection on a density gradient of approximately the complete density range of rat lung ATII cells. Subsequently, to exclude contaminating macrophages and lymphocytes from this fraction, the authors have exploited the fact that the contaminating cells have leukocyte common antigen (LC) on their surfaces, whereas the ATII cells do not. Our procedure yields 2 X 10(7) ATII cells per rat in a fraction which is 90% pure; the cells are immediately available for biochemical or pharmacologic analysis and represent a 90-95% recovery of the ATII cells loaded onto the density gradient. The cells retain their in vivo morphologic characteristics, including their polarity.
Full text
PDF








Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- AVERY M. E., MEAD J. Surface properties in relation to atelectasis and hyaline membrane disease. AMA J Dis Child. 1959 May;97(5 Pt 1):517–523. doi: 10.1001/archpedi.1959.02070010519001. [DOI] [PubMed] [Google Scholar]
- Adamson I. Y., Bowden D. H. The type 2 cell as progenitor of alveolar epithelial regeneration. A cytodynamic study in mice after exposure to oxygen. Lab Invest. 1974 Jan;30(1):35–42. [PubMed] [Google Scholar]
- Bachofen M., Weibel E. R. Basic pattern of tissue repair in human lungs following unspecific injury. Chest. 1974 Apr;65(Suppl):14S–19S. doi: 10.1378/chest.65.4_supplement.14s. [DOI] [PubMed] [Google Scholar]
- Brown S. E., Goodman B. E., Crandall E. D. Type II alveolar epithelial cells in suspension: separation by density and velocity. Lung. 1984;162(5):271–280. doi: 10.1007/BF02715657. [DOI] [PubMed] [Google Scholar]
- CLEMENTS J. A. Surface phenomena in relation to pulmonary function. Physiologist. 1962 Feb;5:11–28. [PubMed] [Google Scholar]
- Clements J. A. Functions of the alveolar lining. Am Rev Respir Dis. 1977 Jun;115(6 Pt 2):67–71. doi: 10.1164/arrd.1977.115.S.67. [DOI] [PubMed] [Google Scholar]
- Dalchau R., Kirkley J., Fabre J. W. Monoclonal antibody to a human leukocyte-specific membrane glycoprotein probably homologous to the leukocyte-common (L-C) antigen of the rat. Eur J Immunol. 1980 Oct;10(10):737–744. doi: 10.1002/eji.1830101003. [DOI] [PubMed] [Google Scholar]
- Darrah H. K., Hedley-Whyte J. Rapid incorporation of palmitate into lung: site and metabolic fate. J Appl Physiol. 1973 Feb;34(2):205–213. doi: 10.1152/jappl.1973.34.2.205. [DOI] [PubMed] [Google Scholar]
- Dobbs L. G., Geppert E. F., Williams M. C., Greenleaf R. D., Mason R. J. Metabolic properties and ultrastructure of alveolar type II cells isolated with elastase. Biochim Biophys Acta. 1980 Jun 23;618(3):510–523. doi: 10.1016/0005-2760(80)90270-2. [DOI] [PubMed] [Google Scholar]
- Evans M. J., Cabral L. J., Stephens R. J., Freeman G. Renewal of alveolar epithelium in the rat following exposure to NO2. Am J Pathol. 1973 Feb;70(2):175–198. [PMC free article] [PubMed] [Google Scholar]
- Fabre J. W., Williams A. F. Quantitative serological analysis of a rabbit anti-rat lymphocyte serum and preliminary biochemical characterisation of the major antigen recognised. Transplantation. 1977 Apr;23(4):349–359. doi: 10.1097/00007890-197704000-00009. [DOI] [PubMed] [Google Scholar]
- Farrell P. M., Avery M. E. Hyaline membrane disease. Am Rev Respir Dis. 1975 May;111(5):657–688. doi: 10.1164/arrd.1975.111.5.657. [DOI] [PubMed] [Google Scholar]
- Fisher A. B., Furia L., Berman H. Metabolism of rat granular pneumocytes isolated in primary culture. J Appl Physiol Respir Environ Exerc Physiol. 1980 Oct;49(4):743–750. doi: 10.1152/jappl.1980.49.4.743. [DOI] [PubMed] [Google Scholar]
- Gil J., Reiss O. K. Isolation and characterization of lamellar bodies and tubular myelin from rat lung homogenates. J Cell Biol. 1973 Jul;58(1):152–171. doi: 10.1083/jcb.58.1.152. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Goerke J. Lung surfactant. Biochim Biophys Acta. 1974 Dec 16;344(3-4):241–261. doi: 10.1016/0304-4157(74)90009-4. [DOI] [PubMed] [Google Scholar]
- Goodman B. E., Crandall E. D. Dome formation in primary cultured monolayers of alveolar epithelial cells. Am J Physiol. 1982 Jul;243(1):C96–100. doi: 10.1152/ajpcell.1982.243.1.C96. [DOI] [PubMed] [Google Scholar]
- Greenleaf R. D., Mason R. J., Williams M. C. Isolation of alveolar type II cells by centrifugal elutriation. In Vitro. 1979 Sep;15(9):673–684. doi: 10.1007/BF02618246. [DOI] [PubMed] [Google Scholar]
- Hirsch J. G., Fedorko M. E. Ultrastructure of human leukocytes after simultaneous fixation with glutaraldehyde and osmium tetroxide and "postfixation" in uranyl acetate. J Cell Biol. 1968 Sep;38(3):615–627. doi: 10.1083/jcb.38.3.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kikkawa Y., Yoneda K., Smith F., Packard B., Suzuki K. The type II epithelial cells of the lung. II. Chemical composition and phospholipid synthesis. Lab Invest. 1975 Mar;32(3):295–302. [PubMed] [Google Scholar]
- Kikkawa Y., Yoneda K. The type II epithelial cell of the lung. I. Method of isolation. Lab Invest. 1974 Jan;30(1):76–84. [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Mason R. J., Williams M. C., Greenleaf R. D., Clements J. A. Isolation and properties of type II alveolar cells from rat lung. Am Rev Respir Dis. 1977 Jun;115(6):1015–1026. doi: 10.1164/arrd.1977.115.6.1015. [DOI] [PubMed] [Google Scholar]
- Mettler N. R., Gray M. E., Schuffman S., LeQuire V. S. beta-Adrenergic induced synthesis and secretion of phosphatidylcholine by isolated pulmonary alveolar type II cells. Lab Invest. 1981 Dec;45(6):575–586. [PubMed] [Google Scholar]
- Meyer K. R., Witschi H., Ullrich R. L. Proliferative responses of type 2 lung epithelial cells after X rays and fission neutrons. Radiat Res. 1980 Jun;82(3):559–569. [PubMed] [Google Scholar]
- O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
- Pochron S. F., Nardone L. L., Kikkawa Y. The effect of anti-oxidants and medium composition on isolation and culture of alveolar type II pneumocytes. Cell Biol Int Rep. 1983 Nov;7(11):963–970. doi: 10.1016/0309-1651(83)90216-3. [DOI] [PubMed] [Google Scholar]
- Skillrud D. M., Martin W. J., 2nd The isolation of rat alveolar type II cells: a simplified approach using Percoll density centrifugation. Lung. 1984;162(4):245–252. doi: 10.1007/BF02715652. [DOI] [PubMed] [Google Scholar]
- Smith F. B., Kikkawa Y. The type II epithelial cells of the lung. V. Synthesis of phosphatidyl glycerol in isolated type II cells and pulmonary alveolar macrophages. Lab Invest. 1979 Feb;40(2):172–177. [PubMed] [Google Scholar]
- Wysocki L. J., Sato V. L. "Panning" for lymphocytes: a method for cell selection. Proc Natl Acad Sci U S A. 1978 Jun;75(6):2844–2848. doi: 10.1073/pnas.75.6.2844. [DOI] [PMC free article] [PubMed] [Google Scholar]



