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Journal of Lipid Research, Vol. 46, 93-103, January 2005
Copyright © 2005 by American Society for Biochemistry and Molecular Biology

Characterization of differentiated subcutaneous and visceral adipose tissue from children

: the influences of TNF- and IGF-I

Malcolm Grohmann^1,*, Matthew Sabin, Jeff Holly^*, Julian Shield, Elizabeth Crowne^2, and Claire Stewart^2,*

^* Department of Surgery, University of Bristol, Bristol Royal Infirmary, Bristol BS2 8HW, United Kingdom
Department of Pediatric Endocrinology, Institute of Child Health, Royal Hospital for Children, Bristol BS2 8AE, United Kingdom

¹ To whom correspondence should be addressed. e-mail: 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, characterize, and compare primary cell cultures of subcutaneous and visceral preadipocytes from 16 normal prepubertal children (10 male and 6 female). Subculture techniques were developed to increase cell number and allow differentiation using a chemically defined serum-free medium. Removal of insulin from the differentiation medium prevented adipogenesis in both subcutaneous and visceral preadipocytes, whereas coincubation with rosiglitazone markedly enhanced glycerol-3-phosphate dehydrogenase activity, peroxisome proliferator-activated receptor 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 compared with visceral cells. Downregulation and reorganization of the cytoskeleton appeared comparable. We further demonstrate regional differences in adipogenesis manipulation. Tumor necrosis factor- was more effective at inhibiting differentiation in subcutaneous cells, whereas insulin-like growth factor-I stimulated differentiation more effectively in visceral cells. Insulin-like growth factor binding protein-3 enhanced differentiation equally.

These observations may have important physiological and pharmacological implications for the development of obesity in later life.

Abbreviations: BMI, body mass index; GPDH, glycerol-3-phosphate dehydrogenase; IGF-I, insulin-like growth factor-I; IGFBP-3, insulin-like growth factor binding protein-3; ORO, Oil Red O; PPAR, peroxisome proliferator-activated receptor ; TNF-, tumor necrosis factor-

Supplementary key words adipocyte • glycerol-3-phosphate dehydrogenase • peroxisome proliferator-activated receptor • actin • tumor necrosis factor • insulin-like growth factor-I • insulin-like growth factor binding protein-3 • adiponectin

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