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A more recent version of this article appeared on January 1, 2005

Papers In Press, published online ahead of print October 16, 2004
J. Lipid Res., doi:10.1194/jlr.M400295-JLR200
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Submitted on August 3, 2004
Revised on October 6, 2004
Accepted on October 12, 2004

Characterisation of a physiologically relevant model to investigate the differentiation of children's subcutaneous and visceral adipose tissue and the influences of TNFalpha and IGF-I

Malcolm J. Grohmann, Matthew A. Sabin, Jeff M. Holly, Julian P. Shield, Elizabeth C. Crowne, and Claire E. Stewart

Department of Surgery, University of Bristol, Bristol BS2 8HW

Corresponding Author: m.j.grohmann{at}bristol.ac.uk

The relationship between subcutaneous and visceral adipocyte metabolism and development has been extensively studied in adult but not in pediatric tissue. Our aim was to isolate, develop, characterise and compare primary cell cultures of subcutaneous and visceral preadipocytes from 16 normal prepubertal children (10 male and 6 female). Sub-culture techniques were developed to increase cell number and allow differentiation using a chemically defined serum-free media. Removal of insulin from the differentiation media prevented adipogenesis in both subcutaneous and visceral preadipocytes, while a co-incubation with rosiglitazone markedly enhanced Glycerol-3-phosphate dehydrogenase (GPDH) activity, peroxisome proliferator activated receptor-gamma (PPARgamma ) expression, and triglyceride accumulation in cells from both fat depots. Adiponectin secretion increased with differentiation from undetectable levels at day 0. Histological analyses demonstrated significant differences in lipid droplet number and size, with subcutaneous cells having fewer but larger vesicles when compared with visceral. Down-regulation and re-organisation of the cytoskeleton appeared comparable. We further demonstrate regional differences in adipogenesis manipulation. Tumour necrosis factor-alpha (TNFalpha )was more effective at inhibiting differentiation in subcutaneous cells, whereas IGF-I stimulated differentiation more effectively in visceral cells. IGFBP-3 enhanced differentiation equally. These observations may have important physiological and pharmacological implications for the development of obesity in later life.


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