HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: D500037-JLR200v1 47/2/450    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Wolins, N. E. Articles by Bickel, P. E. Search for Related Content PubMed PubMed Citation Articles by Wolins, N. E. Articles by Bickel, P. E. Social Bookmarking                      What's this?

Journal of Lipid Research, Vol. 47, 450-460, February 2006
Copyright © 2006 by American Society for Biochemistry and Molecular Biology

Methods

OP9 mouse stromal cells rapidly differentiate into adipocytes: characterization of a useful new model of adipogenesis

Nathan E. Wolins^*, Benjamin K. Quaynor^*, James R. Skinner^*, Anatoly Tzekov^*, Changwon Park, Kyunghee Choi and Perry E. Bickel^1,*,

^* Division of Endocrinology, Metabolism, and Lipid Research, Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110
Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, MO 63110
Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO 63110

Published, JLR Papers in Press, November 30, 2005

¹ To whom correspondence should be addressed. e-mail: pbickel{at}im.wustl.edu

Much knowledge of adipocyte biology has been learned from cell culture models, most notably 3T3-L1 cells. The 3T3-L1 model has several limitations, including the requirement of 2 weeks to generate adipocytes and the waning of adipogenic potential in culture. We have investigated the capacity of OP9 cells, a line of bone marrow-derived mouse stromal cells, to recapitulate adipogenesis. When OP9 cells are given any one of three adipogenic stimuli, they rapidly accumulate triacylglycerol, assume adipocyte morphology, and express adipocyte late marker proteins, including glucose transporter 4 and adiponectin. OP9 cells can differentiate into adipocytes within 2 days. This rapid rate of differentiation allows for the detection of transiently expressed proteins in mature OP9 adipocytes. Adipogenesis in OP9 cells involves the master transcriptional regulator of adipocyte differentiation, peroxisome proliferator-activated receptor (PPAR). OP9 cells are late preadipocytes in that, before the addition of adipogenic stimuli, they express the adipocyte proteins CCAAT/enhancer binding proteins and ß, PPAR, sterol-regulatory element binding protein-1, S3-12, and perilipin. OP9 differentiation is not diminished by maintenance in culture at high cell density or by long periods in continuous culture, thereby facilitating the generation of stable cell lines that retain adipogenic potential. Thus, the unique features of OP9 cells will expedite the study of adipocyte biology.

Abbreviations: ADC, adipogenic cocktail; BMP-4, bone morphogenic protein 4; C/EBP, CCAAT/enhancer binding protein; DN, dominant negative; eGFP-perilipin, perilipin tagged at its N terminus with enhanced green fluorescent protein; GLUT4, glucose transporter 4; IO, insulin oleate; M-CSF, macrophage colony-stimulating factor; PPAR, peroxisome proliferator-activated receptor ; SR, serum replacement; SREBP, sterol-regulatory element binding protein; SSC, side scatter; TAG, triacylglycerol

Supplementary key words bone marrow • flow cytometry • transfection • triacylglycerol • glucose transporter 4 • glucose uptake • peroxisome proliferator-activated receptor • S3-12 • perilipin • 3T3-L1

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				A. C. Doran, N. Meller, A. Cutchins, H. Deliri, R. P. Slayton, S. N. Oldham, J. B. Kim, S. R. Keller, and C. A. McNamara The Helix-Loop-Helix Factors Id3 and E47 Are Novel Regulators of Adiponectin Circ. Res., September 12, 2008; 103(6): 624 - 634. [Abstract] [Full Text] [PDF]

				M. J. Carper, W. T. Cade, M. Cam, S. Zhang, A. Shalev, K. E. Yarasheski, and S. Ramanadham HIV-protease inhibitors induce expression of suppressor of cytokine signaling-1 in insulin-sensitive tissues and promote insulin resistance and type 2 diabetes mellitus Am J Physiol Endocrinol Metab, March 1, 2008; 294(3): E558 - E567. [Abstract] [Full Text] [PDF]

				P. C. Kienesberger, A. Lass, K. Preiss-Landl, H. Wolinski, S. D. Kohlwein, R. Zimmermann, and R. Zechner Identification of an Insulin-regulated Lysophospholipase with Homology to Neuropathy Target Esterase J. Biol. Chem., February 29, 2008; 283(9): 5908 - 5917. [Abstract] [Full Text] [PDF]

				D. Li, Q. Kang, and D.-M. Wang Constitutive Coactivator of Peroxisome Proliferator-Activated Receptor (PPAR{gamma}), a Novel Coactivator of PPAR{gamma} that Promotes Adipogenesis Mol. Endocrinol., October 1, 2007; 21(10): 2320 - 2333. [Abstract] [Full Text] [PDF]

				N. E. Wolins, B. K. Quaynor, J. R. Skinner, A. Tzekov, M. A. Croce, M. C. Gropler, V. Varma, A. Yao-Borengasser, N. Rasouli, P. A. Kern, et al. OXPAT/PAT-1 Is a PPAR-Induced Lipid Droplet Protein That Promotes Fatty Acid Utilization Diabetes, December 1, 2006; 55(12): 3418 - 3428. [Abstract] [Full Text] [PDF]

				S. E. Gale, A. Frolov, X. Han, P. E. Bickel, L. Cao, A. Bowcock, J. E. Schaffer, and D. S. Ory A Regulatory Role for 1-Acylglycerol-3-phosphate-O-acyltransferase 2 in Adipocyte Differentiation J. Biol. Chem., April 21, 2006; 281(16): 11082 - 11089. [Abstract] [Full Text] [PDF]