title: 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 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.
authors.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 chrisna.durandt@up.ac.za.
authors.forename: Chrisna
authors.initials: C
authors.lastname: Durandt
authors.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.
authors.forename: Fiona A
authors.initials: FA
authors.lastname: van Vollenstee
authors.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.
authors.forename: Carla
authors.initials: C
authors.lastname: Dessels
authors.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.
authors.forename: Karlien
authors.initials: K
authors.lastname: Kallmeyer
authors.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.
authors.forename: Danielle
authors.initials: D
authors.lastname: de Villiers
authors.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.
authors.forename: Candice
authors.initials: C
authors.lastname: Murdoch
authors.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.
authors.forename: Marnie
authors.initials: M
authors.lastname: Potgieter
authors.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.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Pepper
mesh_terms: D017667:Adipocytes / Q000166:cytology* / Q000378:metabolism; D050156:Adipogenesis; D022763:CCAAT-Enhancer-Binding Protein-alpha / Q000235:genetics; D018955:CD36 Antigens / Q000235:genetics / Q000378:metabolism; D050556:Fatty Acid-Binding Proteins / Q000235:genetics; D005434:Flow Cytometry / Q000379:methods*; D005786:Gene Expression Regulation; D006801:Humans; D008856:Microscopy, Fluorescence; D047495:PPAR gamma / Q000235:genetics
chemical_list: D022763:CCAAT-Enhancer-Binding Protein-alpha; D018955:CD36 Antigens; C493410:FABP4 protein, human; D050556:Fatty Acid-Binding Proteins; D047495:PPAR gamma
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
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