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Fat Depot–Specific Characteristics Are Retained in Strains Derived From Single Human Preadipocytes

¹ Department of Medicine, Boston University, Boston, Massachusetts
² AdipoGenix, Boston, Massachusetts
³ Department of Surgery, Creighton University, Omaha, Nebraska
⁴ Henry Wellcome Biogerontology Laboratory, Institute for Ageing and Health, University of Newcastle, Newcastle, U.K
⁵ Department of Biochemistry, Boston University, Boston, Massachusetts

Address correspondence and reprint requests to James L. Kirkland, MD, PhD, Boston University, 88 East Newton St., Robinson 2, Boston, MA 02118. E-mail: kirkland{at}bu.edu

Abbreviations: C/EBP, CCAAT/enhancer-binding protein ; FBS, fetal bovine serum; G3PD, glycerol-3-phosphate dehydrogenase; hTERT, human telomere reverse transcriptase; mRNA, messenger RNA; PPAR, peroxisome proliferator–activated receptor; TNF, tumor necrosis factor

Fat depots vary in size, function, and potential contribution to disease. Since fat tissue turns over throughout life, preadipocyte characteristics could contribute to this regional variation. To address whether preadipocytes from different depots are distinct, we produced preadipocyte strains from single abdominal subcutaneous, mesenteric, and omental human preadipocytes by stably expressing human telomere reverse transcriptase (hTERT). These strains could be subcultured repeatedly and retained capacity for differentiation, while primary preadipocyte adipogenesis and replication declined with subculturing. Primary omental preadipocytes, in which telomeres were longest, replicated more slowly than mesenteric or abdominal subcutaneous preadipocytes. Even after 40 population doublings, replication, abundance of the rapidly replicating preadipocyte subtype, and resistance to tumor necrosis factor –induced apoptosis were highest in subcutaneous, intermediate in mesenteric, and lowest in omental hTERT-expressing strains, as in primary preadipocytes. Subcutaneous hTERT-expressing strains accumulated more lipid and expressed more adipocyte fatty acid–binding protein (aP2), peroxisome proliferator–activated receptor 2, and CCAAT/enhancer-binding protein than omental cells, as in primary preadipocytes, while hTERT abundance was similar. Thus, despite dividing 40 population doublings, hTERT strains derived from single preadipocytes retained fat depot–specific cell dynamic characteristics, consistent with heritable processes contributing to regional variation in fat tissue function.

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