AJP - Lung Journal of Applied Physiology
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Am J Physiol Lung Cell Mol Physiol 260: L395-L402, 1991;
1040-0605/91 $5.00
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AJP - Lung Cellular and Molecular Physiology, Vol 260, Issue 6 395-L402, Copyright © 1991 by American Physiological Society

ARTICLES

Limited division of low-density adult rat type II pneumocytes in serum-free culture

A. K. Tanswell, P. J. Byrne, R. N. Han, J. D. Edelson and V. K. Han
Department of Medicine, St. Michael's Hospital, University of Toronto, Ontario, Canada.

A previously reported serum-free medium for the culture of adult rat type II pneumocytes has been further defined by the addition of 10 mM N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid buffer and 700 U/ml catalase. Both additions significantly enhanced [3H]thymidine incorporation into pneumocyte DNA between days 5 and 6 of culture. In the presence of 100 micrograms/ml bovine pituitary extract, both insulin-like growth factor (IGF) I (20 ng/ml) and II (25 ng/ml) further increased [3H]thymidine incorporation into pneumocyte DNA over that observed with the basal medium. When type II pneumocytes were plated at 2.75 X 10(5) cells/cm2 there was a plating efficiency of 50 +/- 5%, and the increased DNA synthesis in response to IGFs was not associated with an increase in cell number. When the plating number was reduced to 1.6 X 10(5) cells/cm2 the plating efficiency was only 23 +/- 2%. At this low cell density a doubling of cell number was observed in response to IGF-I between days 4 and 9 of culture.


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