Heterogeneity in the physiological states and pharmacological responses of differentiating 3T3-L1 preadipocytes

J Cell Biol. 2009 Nov 2;187(3):375-84. doi: 10.1083/jcb.200904140. Epub 2009 Oct 26.


Increases in key components of adipogenesis and lipolysis pathways correlate at the population-averaged level during adipogenesis. However, differentiating preadipocytes are highly heterogeneous in cellular and lipid droplet (LD) morphologies, and the degree to which individual cells follow population-averaged trends is unclear. In this study, we analyze the molecular heterogeneity of differentiating 3T3-L1 preadipocytes using immunofluorescence microscopy. Unexpectedly, we only observe a small percentage of cells with high simultaneous expression of markers for adipogenesis (peroxisome proliferator-activated receptor gamma [PPARgamma], CCAAT/enhancer-binding protein alpha, and adiponectin) and lipid accumulation (hormone-sensitive lipase, perilipin A, and LDs). Instead, we identify subpopulations of cells with negatively correlated expressions of these readouts. Acute perturbation of adipocyte differentiation with PPARgamma agonists, forskolin, and fatty acids induced subpopulation-specific effects, including redistribution of the percentage of cells in observed subpopulations and differential expression levels of PPARgamma. Collectively, our results suggested that heterogeneity observed during 3T3-L1 adipogenesis reflects a dynamic mixture of subpopulations with distinct physiological states.

Publication types

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

MeSH terms

  • Adipocytes / cytology
  • Adipocytes / drug effects
  • Adipogenesis / physiology
  • Adiponectin / metabolism
  • Animals
  • CCAAT-Binding Factor / metabolism
  • Cell Differentiation / drug effects*
  • Colforsin / pharmacology
  • Fatty Acids / pharmacology
  • Genetic Markers
  • Lipid Metabolism
  • Mice
  • Microscopy, Fluorescence
  • NIH 3T3 Cells
  • PPAR gamma / agonists
  • PPAR gamma / metabolism
  • Time Factors


  • Adiponectin
  • CCAAT-Binding Factor
  • Fatty Acids
  • Genetic Markers
  • PPAR gamma
  • Colforsin