J. Anim Sci.
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J. Anim Sci. 1988. 66:3267-3278.
© 1988 American Society of Animal Science
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The Effect of Insulin on Primary Cultures of Rat Preadipocytes Grown in Fetal or Postnatal Pig Serum

G. J. Hausman1,2,4, and D. E. Jewell3

Abstract

Stromal vascular cell cultures, prepared from the inguinal pads of 50-g Sprague Dawley rats, were exposed to media with 10% fetal pig serum which is inherently low in insulin, for the first 3 to 5 d of culture. Insulin was supplemented to media for periods of 2 to 6 d. In cultures treated (2 to 4 d) with 10–9, 10–10 or 10–11 M insulin, differentiated cells (lipid and esterase staining) appeared 1.5 to 2 times wider than differentiated cells in control cultures. At 10–9 M insulin (4 to 5 d), in cultures grown in the presence of fetal pig serum the number of esterase reactive cells was increased twofold to threefold. The percentage of total cells that were esterase reactive was elevated 50 to 300% relative to control cultures. Insulin-treated preadipocytes were more reactive for lipoprotein lipase activity (histochemical assay) compared with reactivity of control cells. Quantitative analysis of percentage of light transmittance (Zeiss photometer) through stained cells indicated an increase (P < .001) in lipoprotein lipase staining at 10–9, 10–11 and 10–13 M insulin (2 d). The specific activity of glycerol phosphate dehydrogenase was elevated twofold to threefold (P < .05) and soluble protein elevated 50 to 100% (P < .05) in cultures treated (3 to 6 d) with 10–9 M insulin. Decreasing the cell plating density (50%) in cultures grown in the presence of pig serum reduced the elevation in enzyme activity induced by insulin in preadipocyte cultures. Physiological levels of insulin enhanced lipogenic enzyme activity in preadipocytes and may enhance the conversion of stromal cells to preadipocytes.

Footnotes

1 The authors thank Glenda B. Thomas for the able execution of these studies, and thanks are expressed to R. W. Seerley, Dept. of Anim. Sci., Univ. of Georgia, for providing animal facilities and for maintaining the animals used in this study.

2 Mention of a trade name, proprietary product, or specific equipment does not constitute a guarantee or warranty by the USDA and does not imply its approval to the exclusion of other products that may be suitable.

3 Purina Mills Inc., 1401 S. Hanley Road, St. Louis, MO 63166.

4 Richard B. Russell Res. Center, Animal Physiology Res. Unit. USDA-ARS, P. O. Box 5677, Athens, Georgia 30613.






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Copyright © 1988 by the American Society of Animal Science.