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Papers In Press, published online ahead of print March 1, 2008
J. Lipid Res., doi:10.1194/jlr.D700029-JLR200

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Journal of Lipid Research, Vol. 49, 679-685, March 2008
Copyright © 2008 by American Society for Biochemistry and Molecular Biology

Methods

Conditionally immortalized white preadipocytes: a novel adipocyte model

D. L. Morganstein^*, M. Christian^*, J. J. O. Turner, M. G. Parker^* and R. White^1,*

^* Institute of Reproductive and Developmental Biology, Imperial College London, London W12 0NN, United Kingdom
Institute of Metabolic Medicine, Imperial College London, London W12 0NN, United Kingdom

The online version of this article (available at http://www.jlr.org) contains supplementary data in the form of three figures.

Published, JLR Papers in Press, November 28, 2007.

¹ To whom correspondence should be addressed. e-mail: roger.white{at}imperial.ac.uk

This study describes a novel approach to generate conditionally immortalized preadipocyte cell lines from white adipose tissue (IMWAT) that can be induced to differentiate into white adipocytes even after expansion in culture. Such adipocytes express markers of white fat such as peroxisome proliferator-activated receptor and aP2 but not brown fat markers, have an intact insulin signaling pathway, and express proinflammatory cytokines. They can be readily transduced with adenoviral vectors, allowing them to be used to investigate the consequences of the depletion of specific adipocyte factors using short hairpin RNA. This approach has been used to study the effect of reduced expression of the nuclear receptor corepressor receptor interacting protein 140 (RIP140), a regulator of adipocyte function. The depletion of RIP140 results in changes in metabolic gene expression that resemble those in adipose tissue of the RIP140 null mouse. Thus, IMWAT cells provide a novel model for adipocytes that are derived from preadipocytes rather than fibroblasts and provide an alternative system to primary preadipocytes for the investigation of adipocyte function.

Supplementary key words immortalization • adenovirus • short hairpin RNA • receptor interacting protein 140

Abbreviations: GFP, Green Fluorescent Protein; IL-6, interleukin 6; IMBAT, immortalized brown adipocyte; IMWAT, immortalized white adipocyte; LPS, lipopolysaccharide; PRDM, PR domain containing 16; shRNA, short hairpin RNA; Rb, Retinoblastoma; TNF-, tumor necrosis factor-; Ucp1, Uncoupling Protein 1

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