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

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 w...

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Bibliographic Details
Published in:Journal of lipid research 2016-04, Vol.57 (4), p.729-742
Main Authors: Durandt, Chrisna, van Vollenstee, Fiona A., Dessels, Carla, Kallmeyer, Karlien, de Villiers, Danielle, Murdoch, Candice, Potgieter, Marnie, Pepper, Michael S.
Format: Article
Language:English
Subjects:
adipocyte differentiation
Adipocytes - cytology
Adipocytes - metabolism
Adipogenesis
adipose tissue-derived stromal cells
Bodipy 493/503
CCAAT-Enhancer-Binding Protein-alpha - genetics
CD36 Antigens - genetics
CD36 Antigens - metabolism
cluster of differentiation 36
Fatty Acid-Binding Proteins - genetics
fatty/acid binding protein
Flow Cytometry - methods
gene expression
Gene Expression Regulation
Humans
lipid droplet
Methods
Microscopy, Fluorescence
Nile Red
PPAR gamma - genetics
triglyceride
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Summary: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.
ISSN:0022-2275
1539-7262
DOI:10.1194/jlr.D065664