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Originally published In Press as doi:10.1194/jlr.D600005-JLR200 on May 4, 2006

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Journal of Lipid Research, Vol. 47, 1859-1864, August 2006
Copyright © 2006 by American Society for Biochemistry and Molecular Biology

Methods

Optimized conditions for measuring lipolysis in murine primary adipocytes

Srikant Viswanadha and Constantine Londos1

Laboratory of Cellular and Developmental Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-8028

Published, JLR Papers in Press, May 4, 2006.

1 To whom correspondence should be addressed. e-mail: deanl{at}intra.niddk.nih.gov

The current literature on lipolysis in murine primary adipocytes is rife with experiments performed under conditions not optimized for reproducible and reliable results. Here, we present conditions for optimizing the measurement of lipolysis in murine adipocytes. We demonstrate that adenosine management is of paramount importance in evaluating the lipolytic response under basal and stimulated conditions. Also, adipocyte concentrations in the 10,000–15,000 cells per milliliter range produce a greater increase in stimulated lipolysis than higher concentrations, and the response is further enhanced by agitating the cells.

Supplementary key words glycerol • adenosine • cell concentrations


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