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

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 adiposederived stromal cells that express CD36 at intermediate/high levels and show that combining CD36 cell surface staining with neutral lipidspecific staining allows us to monitor differentiation of adiposederived stromal cells that express CD36 during adipocyte differentiation in vitro. The gradual increase of CD36 NR cells during the 21 day adipogenesis induction period correlated with upregulation of adipogenesisassociated gene expression.