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Am J Physiol Regul Integr Comp Physiol 258: R198-R204, 1990;
0363-6119/90 $5.00
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AJP - Regulatory, Integrative and Comparative Physiology, Vol 258, Issue 1 198-R204, Copyright © 1990 by American Physiological Society

ARTICLES

Counteracting effects of urea and betaine in mammalian cells in culture

P. H. Yancey and M. B. Burg
Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung, and Blood Institute, Bethesda, Maryland 20892.

Urea and methylamines, such as betaine, are among the major organic osmotic effectors accumulated by organisms under hyperosmotic (high NaCl) stress; the mammalian renal medulla also accumulates such compounds in antidiuresis. Studies on isolated proteins show that urea generally destabilizes protein structure, whereas methylamines are generally stabilizers capable of offsetting the effects of urea. The counteracting-osmolytes hypothesis predicts that cells exposed to high urea concentrations require methylamines for optimal function. In this study, urea, betaine, and other solutes (NaCl, glycerol, sorbitol) were added to growth medium of cultured mammalian cells under conditions in which most solutes entered the cells. For two renal [Madin-Darby canine kidney (MDCK) and PAP-HT25] and one nonrenal (Chinese hamster ovary) cell line, urea (greater than 100 mM) or betaine (greater than 50-100 mM) alone inhibited cell growth and survival, measured as colony-forming efficiency. However, the addition of betaine (up to 120 mM) to media with urea (50-300 mM) greatly increased colony-forming efficiency above that with urea alone. A similar, although less marked effect, was seen on colony sizes with MDCK cells. These results support the counteracting-osmolytes hypothesis.


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