The ability of mesenchymal stromal cells (MSCs) to differentiate into adipocytes provides a cellular model ofhuman origin to study adipogenesis in vitro. One of the major challenges in studying adipogenesis is the lackof tools to identify and monitor the differentiation of various subpopulations within the heterogeneous poolof MSCs. Cluster of differentiation (CD)36 plays an important role in the formation of intracellular lipiddroplets, a key characteristic of adipocyte differentiation/maturation. The objective of this study was todevelop a reproducible quantitative method to study adipocyte differentiation by comparing two lipophilicdyes [Nile Red (NR) and Bodipy 493/503] in combination with CD36 surface marker staining. We identifieda subpopulation of adiposederivedstromal cells that express CD36 at intermediate/high levels and show thatcombining CD36 cell surface staining with neutral lipidspecificstaining allows us to monitor differentiationof adiposederivedstromal cells that express CD36 during adipocyte differentiation in vitro.The gradual increase of CD36 NR cells during the 21 day adipogenesis inductionperiod correlated with upregulation of adipogenesisassociatedgene expression.
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