Abstract
Flow cytometry and fluorescence-activated cell sorting (FACS) techniques have significantly advanced the characterization of adipocyte precursor cell (APC) populations. They allow immunophenotyping, quantification, and isolation of distinct populations, which is critical for understanding adipose tissue development and homeostasis. Here, we describe the identification and purification of adipocyte precursor cells using flow cytometry and FACS, defined by previously established surface marker profiles. In addition, we describe the mouse models and whole adipose tissue visualization techniques that will enable us to characterize the plasticity and the cellular origin of APCs.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Park A, Kim WK, Bae KH (2014) Distinction of white, beige and brown adipocytes derived from mesenchymal stem cells. World J Stem Cells 6(1):33–42
Harms M, Seale P (2013) Brown and beige fat: development, function and therapeutic potential. Nat Med 19(10):1252–1263
Moulin K et al (2001) Emergence during development of the white-adipocyte cell phenotype is independent of the brown-adipocyte cell phenotype. Biochem J 356(Pt 2):659–664
Uhm M, Saltiel AR (2015) White, brown, and beige; type 2 immunity gets hot. Immunity 42(1):15–17
Bostrom P et al (2012) A PGC1-alpha-dependent myokine that drives brown-fat-like development of white fat and thermogenesis. Nature 481(7382):463–468
Xu X et al (2011) Exercise ameliorates high-fat diet-induced metabolic and vascular dysfunction, and increases adipocyte progenitor cell population in brown adipose tissue. Am J Physiol Regul Integr Comp Physiol 300(5):R1115–R1125
Rosenwald M, Wolfrum C (2014) The origin and definition of brite versus white and classical brown adipocytes. Adipocytes 3(1):4–9
Rosen ED, MacDougald OA (2006) Adipocyte differentiation from the inside out. Nat Rev Mol Cell Biol 7(12):885–896
Wu J et al (2012) Beige adipocytes are a distinct type of thermogenic fat cell in mouse and human. Cell 150(2):366–376
Hui X et al (2015) Adiponectin enhances cold-induced Browning of subcutaneous adipose tissue via promoting M2 macrophage proliferation. Cell Metab 22(2):279–290
Odegaard JI et al (2016) Perinatal licensing of thermogenesis by IL-33 and ST2. Cell 166(4):841–854
Rosenwald M et al (2013) Bi-directional interconversion of brite and white adipocytes. Nat Cell Biol 15(6):659–667
Feil S, Valtcheva N, Feil R (2009) Inducible Cre mice. Methods Mol Biol 530:343–363
Jeffery E et al (2014) Characterization of Cre recombinase models for the study of adipose tissue. Adipocytes 3(3):206–211
Krueger KC et al (2014) Characterization of Cre recombinase activity for in vivo targeting of adipocyte precursor cells. Stem Cell Reports 3(6):1147–1158
Berry DC, Jiang Y, Graff JM (2016) Mouse strains to study cold-inducible beige progenitors and beige adipocyte formation and function. Nat Commun 7:10184
Kretzschmar K, Watt FM (2012) Lineage tracing. Cell 148(1–2):33–45
Livet J et al (2007) Transgenic strategies for combinatorial expression of fluorescent proteins in the nervous system. Nature 450(7166):56–62
Berry R, Rodeheffer MS (2013) Characterization of the adipocyte cellular lineage in vivo. Nat Cell Biol 15(3):302–308
Muzumdar MD et al (2007) A global double-fluorescent Cre reporter mouse. Genesis 45(9):593–605
Rodeheffer MS, Birsoy K, Friedman JM (2008) Identification of white adipocyte progenitor cells in vivo. Cell 135(2):240–249
Tang W et al (2008) White fat progenitor cells reside in the adipose vasculature. Science 322(5901):583–586
Cao Y (2010) Adipose tissue angiogenesis as a therapeutic target for obesity and metabolic diseases. Nat Rev Drug Discov 9(2):107–115
Tran KV et al (2012) The vascular endothelium of the adipose tissue gives rise to both white and brown fat cells. Cell Metab 15(2):222–229
Gupta RK et al (2012) Zfp423 expression identifies committed preadipocytes and localizes to adipose endothelial and perivascular cells. Cell Metab 15(2):230–239
Acknowledgments
This work was funded by grants from the Natural Sciences and Engineering Research Council (NSERC) of Canada, Pilot and Feasibility Study Grant of Banting & Best Diabetes Centre (BBDC), Centre for Healthy Active Kids (CHAK) Micro-grant and Sickkids Start-up fund to H.-K.S. J.-H.M is supported by the Restracomp fellowship from The Hospital for Sick Children.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Lee, J.H., Yeganeh, A., Konoeda, H., Moon, J.H., Sung, HK. (2018). Flow Cytometry and Lineage Tracing Study for Identification of Adipocyte Precursor Cell (APC) Populations. In: Delgado-Olguin, P. (eds) Mouse Embryogenesis. Methods in Molecular Biology, vol 1752. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7714-7_11
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7714-7_11
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7713-0
Online ISBN: 978-1-4939-7714-7
eBook Packages: Springer Protocols