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Regulation of lipid accumulation in 3T3-L1 cells: insulin-independent and combined effects of fatty acids and insulin

Published online by Cambridge University Press:  01 January 2008

T. A. Kokta ,
A. L. Strat ,
M. R. Papasani ,
J. I. Szasz ,
M. V. Dodson  and
R. A. Hill
T. A. Kokta
Affiliation:
Department of Animal and Veterinary Science, University of Idaho, PO Box 442330, Moscow, ID 83844-2330, USA
A. L. Strat
Affiliation:
Department of Animal and Veterinary Science, University of Idaho, PO Box 442330, Moscow, ID 83844-2330, USA
M. R. Papasani
Affiliation:
Department of Animal and Veterinary Science, University of Idaho, PO Box 442330, Moscow, ID 83844-2330, USA
J. I. Szasz
Affiliation:
Department of Animal and Veterinary Science, University of Idaho, PO Box 442330, Moscow, ID 83844-2330, USA
M. V. Dodson
Affiliation:
Department of Animal Sciences, Washington State University, Pullman, WA 99164, USA
R. A. Hill*
Affiliation:
Department of Animal and Veterinary Science, University of Idaho, PO Box 442330, Moscow, ID 83844-2330, USA
*
E-mail: [email protected]
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Abstract

The insulin-independent and combined effects of fatty acids (FA; linoleic and oleic acids) and insulin in modulating lipid accumulation and adipogenesis in 3T3-L1 cells was investigated using a novel protocol avoiding the effects of a complex hormone ‘induction’ mixture. 3T3-L1 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) plus serum (control) or in DMEM plus either 0.3 mmol/l linoleic or oleic acids with 0.3 mmol/l FA-free bovine serum albumin in the presence or absence of insulin. Cells were cultured for 4 to 8 days and cell number, lipid accumulation, peroxisome proliferator-activated receptor-gamma (PPAR-γ) and glucose transporter 4 (GLUT-4) protein expression were determined. Cell number appeared to be decreased in comparison with control cultures. In both oleic acid and linoleic acid-treated cells, notably in the absence (and presence) of insulin, oil-red O stain-positive cells showed abundant lipid. The percentage of cells showing lipid accumulation was greater in FA-treated cultures compared with control cells grown in DMEM plus serum (P < 0.001). Treatment with both linoleic and oleic acid-containing media evoked higher levels of PPAR-γ than observed in control cultures (P < 0.05). GLUT-4 protein also increased in response to treatment with both linoleic and oleic acid-containing media (P < 0.001). Lipid accumulation in 3T3-L1 cells occurs in response to either oleic or linoleic acids independently of the presence of insulin. Both PPAR-γ and GLUT-4 protein expression were stimulated. Both proteins are considered markers of adipogenesis, and these observations suggest that these cells had entered the physiological state broadly accepted as differentiated. Furthermore, 3T3-L1 cells can be induced to accumulate lipid in a serum-free medium supplemented with FA, without the use of induction protocols using complex hormone mixtures. We have demonstrated a novel model for the study of lipid accumulation that will improve the understanding of adipogenesis in adipocyte lineage cells.

Keywords

adipogenesisfatty acidsGLUT-4insulinPPAR-γ
Type
Full Paper
Information
animal , Volume 2 , Issue 1 , January 2008 , pp. 92 - 99
Copyright
Copyright © The Animal Consortium 2008

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