Novel flow cytometric approach for the detection of adipocyte subpopulations during adipogenesis |
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Authors: | Chrisna Durandt Fiona A. van Vollenstee Carla Dessels Karlien Kallmeyer Danielle de Villiers Candice Murdoch Marnie Potgieter Michael S. Pepper |
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Affiliation: | Institute for Cellular and Molecular Medicine, South African Medical Research Council Extramural Unit for Stem Cell Research and Therapy, Department of Immunology, Faculty of Health Sciences, University of Pretoria, Pretoria, South Africa |
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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 CD36intermediate/high during adipocyte differentiation in vitro. The gradual increase of CD36intermediate/high/NRpositive cells during the 21 day adipogenesis induction period correlated with upregulation of adipogenesis-associated gene expression. |
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Keywords: | adipocyte differentiation adipose tissue-derived stromal cells Bodipy 493/503 cluster of differentiation 36 fatty/acid binding protein gene expression lipid droplet Nile Red triglyceride |
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