Isolation and study of adipocyte precursors.
White adipose tissue (WAT) is a heterogeneous tissue composed of lipid-filled adipocytes and several nonadipocyte cell populations, including endothelial, blood, uncharacterized stromal, and adipocyte precursor cells. Although lipid-filled adipocytes account for the majority of WAT volume and mass, nonadipocyte cell populations have critical roles in WAT maintenance, growth, and function. As mature adipocytes are terminally differentiated postmitotic cells, differentiation of adipocyte precursors is required for hyperplastic WAT growth during development and in obesity. In this chapter, we present methods to separate adipocyte precursor cells from other nonadipocyte cell populations within WAT for analysis by flow cytometry or purification by fluorescence-activated cell sorting. Additionally, we provide methods to study the adipogenic capacity of purified adipocyte precursor cells ex vivo.
Section of Comparative Medicine, Yale University School of Medicine, New Haven, Connecticut, USA; Yale Stem Cell Center, Yale University School of Medicine, New Haven, Connecticut, USA.
Christopher D
CD
Church
Section of Comparative Medicine, Yale University School of Medicine, New Haven, Connecticut, USA; Department of Molecular, Cell, and Developmental Biology, Yale University School of Medicine, New Haven, Connecticut, USA; Yale Stem Cell Center, Yale University School of Medicine, New Haven, Connecticut, USA.
Ryan
R
Berry
Section of Comparative Medicine, Yale University School of Medicine, New Haven, Connecticut, USA; Department of Molecular, Cell, and Developmental Biology, Yale University School of Medicine, New Haven, Connecticut, USA; Yale Stem Cell Center, Yale University School of Medicine, New Haven, Connecticut, USA. Electronic address: matthew.rodeheffer@yale.edu.
Matthew S
MS
Rodeheffer
D041721:3T3-L1 Cells; D017667:Adipocytes / Q000473:pathology*; D050156:Adipogenesis; D052436:Adipose Tissue, White / Q000254:growth & development / Q000473:pathology*; D000818:Animals; D002454:Cell Differentiation / Q000235:genetics; D049109:Cell Proliferation; D004730:Endothelium, Vascular / Q000473:pathology; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D051379:Mice; D009765:Obesity / Q000473:pathology*
Adipocyte precursor; Adipogenesis; Adipose tissue; Preadipocytes; Progenitor; Stem cell; Stromal vascular