Culture of human adipose tissue explants leads to profound alteration of adipocyte gene expression

Horm Metab Res. 2003 Mar;35(3):158-63. doi: 10.1055/s-2003-39070.

Abstract

Primary culture of adipose tissue has often been used to investigate pharmacological and nutritional regulation of adipocyte gene expression. Possible alteration of adipocyte gene expression by primary culture on its own has not been explored in detail. In order to address this issue, explants were prepared from human subcutaneous adipose tissue recovered from plastic surgery and maintained for 0 to 48 h in DMEM supplemented with 10 % serum. At different time points, adipocytes were isolated from the explants by collagenase digestion, and mRNA expression and lipolysis were studied. Culture was associated with an accumulation of tumor necrosis factor-alpha (TNFalpha) in the culture medium, an increase in anaerobic glycolysis, and an increase in the basal lipolysis. In parallel, a rapid and dramatic decrease in the level of mRNA encoding for several adipocyte-specific proteins such as adipocyte lipid-binding protein, hormone-sensitive lipase, lipoprotein lipase, and peroxisome proliferation activating receptor-gamma2 was observed in isolated adipocytes. These downregulations were reminiscent of a dedifferentiation process. In parallel, primary culture was associated with an increase in adipocyte beta-actin, TNFalpha, glucose transporter-1 and hypoxia-induced factor-1alpha mRNAs. Treatment of explants with agents that increase cAMP (isobutylmethylxanthine and forskolin) prevented TNFalpha production and expression and culture-induced alterations of adipocyte gene expression. These data show that primary culture of human adipose tissue explants dramatically alters adipocyte gene expression.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 1-Methyl-3-isobutylxanthine / pharmacology
  • Actins / genetics
  • Adipocytes / chemistry
  • Adipocytes / drug effects
  • Adipocytes / metabolism
  • Adipose Tissue / drug effects
  • Adipose Tissue / metabolism*
  • Adult
  • Carrier Proteins / genetics
  • Colforsin / pharmacology
  • Culture Media / chemistry
  • Culture Techniques*
  • DNA-Binding Proteins / genetics
  • Fatty Acid-Binding Protein 7
  • Fatty Acid-Binding Proteins
  • Female
  • Gene Expression*
  • Glucose Transporter Type 1
  • Glycolysis
  • Humans
  • Hypoxia-Inducible Factor 1
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Lipolysis
  • Lipoprotein Lipase / genetics
  • Middle Aged
  • Monosaccharide Transport Proteins / genetics
  • Neoplasm Proteins*
  • Nuclear Proteins / genetics
  • RNA, Messenger / analysis
  • Receptors, Cytoplasmic and Nuclear / genetics
  • Sterol Esterase / genetics
  • Transcription Factors / genetics
  • Tumor Necrosis Factor-alpha / genetics
  • Tumor Necrosis Factor-alpha / metabolism
  • Tumor Suppressor Proteins*

Substances

  • Actins
  • Carrier Proteins
  • Culture Media
  • DNA-Binding Proteins
  • FABP7 protein, human
  • Fatty Acid-Binding Protein 7
  • Fatty Acid-Binding Proteins
  • Glucose Transporter Type 1
  • HIF1A protein, human
  • Hypoxia-Inducible Factor 1
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • Monosaccharide Transport Proteins
  • Neoplasm Proteins
  • Nuclear Proteins
  • RNA, Messenger
  • Receptors, Cytoplasmic and Nuclear
  • SLC2A1 protein, human
  • Transcription Factors
  • Tumor Necrosis Factor-alpha
  • Tumor Suppressor Proteins
  • Colforsin
  • Sterol Esterase
  • Lipoprotein Lipase
  • 1-Methyl-3-isobutylxanthine