Novel flow cytometric approach for the detection of adipocyte subpopulations during adipogenesis

J Lipid Res. 2016 Apr;57(4):729-42. doi: 10.1194/jlr.D065664. Epub 2016 Feb 1.

Abstract

The ability of mesenchymal stromal cells (MSCs) to differentiate into adipocytes provides a cellular model of human origin to study adipogenesis in vitro. One of the major challenges in studying adipogenesis is the lack of tools to identify and monitor the differentiation of various subpopulations within the heterogeneous pool of MSCs. Cluster of differentiation (CD)36 plays an important role in the formation of intracellular lipid droplets, a key characteristic of adipocyte differentiation/maturation. The objective of this study was to develop a reproducible quantitative method to study adipocyte differentiation by comparing two lipophilic dyes [Nile Red (NR) and Bodipy 493/503] in combination with CD36 surface marker staining. We identified a subpopulation of adipose-derived stromal cells that express CD36 at intermediate/high levels and show that combining CD36 cell surface staining with neutral lipid-specific staining allows us to monitor differentiation of adipose-derived stromal cells that express CD36(intermediate/high)during adipocyte differentiation in vitro. The gradual increase of CD36(intermediate/high/)NR(positive)cells during the 21 day adipogenesis induction period correlated with upregulation of adipogenesis-associated gene expression.

Keywords: Bodipy 493/503; Nile Red; adipocyte differentiation; adipose tissue-derived stromal cells; cluster of differentiation 36; fatty/acid binding protein; gene expression; lipid droplet; triglyceride.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipocytes / cytology*
  • Adipocytes / metabolism
  • Adipogenesis*
  • CCAAT-Enhancer-Binding Protein-alpha / genetics
  • CD36 Antigens / genetics
  • CD36 Antigens / metabolism
  • Fatty Acid-Binding Proteins / genetics
  • Flow Cytometry / methods*
  • Gene Expression Regulation
  • Humans
  • Microscopy, Fluorescence
  • PPAR gamma / genetics

Substances

  • CCAAT-Enhancer-Binding Protein-alpha
  • CD36 Antigens
  • FABP4 protein, human
  • Fatty Acid-Binding Proteins
  • PPAR gamma