Optimal protocol for the differentiation and metabolic analysis of human adipose stromal cells

Methods Enzymol. 2014:538:49-65. doi: 10.1016/B978-0-12-800280-3.00004-9.

Abstract

Obesity is reaching epidemic proportions so there is growing interest in the mechanisms that regulates adipose tissue development and function. Although murine adipose cell lines are useful for many mechanistic studies, primary human adipose stromal cells (ASCs), which can be isolated from distinct adipose depots and cultured in vitro, have clear translational relevance. We describe the methods to isolate, culture, and differentiate human ASCs to adipocytes that respond to physiologically relevant hormones, such as insulin and β-adrenergic agonists. We also describe methods for assaying hormonal effects on glucose transport and lipolysis.

Keywords: Adipocytes; Adipogenesis; Adipose tissue; Glucose transport; Lipolysis; Preadipocyte.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipocytes / cytology*
  • Adipocytes / metabolism
  • Adipogenesis*
  • Adipose Tissue / cytology*
  • Adipose Tissue / metabolism
  • Adrenergic beta-Agonists / metabolism
  • Cell Culture Techniques / methods
  • Cell Separation / methods
  • Cells, Cultured
  • Humans
  • Insulin / metabolism
  • Lipolysis
  • Obesity / metabolism
  • Stromal Cells / cytology*
  • Stromal Cells / metabolism

Substances

  • Adrenergic beta-Agonists
  • Insulin