title: Isolation and Fluorescence-Activated Cell Sorting of Murine WT1-Expressing Adipocyte Precursor Cells.
abstract: The current global obesity epidemic has triggered increased interest in adipose tissue biology. A major area of attention for many is adipose tissue development. A greater understanding of adipocyte ontogeny could be highly beneficial in answering questions about obesity-associated disease. Recent work has shown that a proportion of mature adipocytes in visceral white adipose tissue are derived from Wt1-expressing adipocyte precursor cells. These adipocyte precursor cells reside within the adipose tissue itself, and are a constituent of the stromal vascular fraction (SVF), along with other, non-adipogenic, cell types. Crucially, heterogeneity exists within the adipocyte precursor population, with only a proportion of cells expressing Wt1. Moreover, it appears that this difference in the precursor cells may influence the mature adipocytes, with Wt1-lineage-positive adipocytes having fewer, larger lipid droplets than the Wt1-lineage negative. Using fluorescence-activated cell sorting, based on specific marker profiles, it is possible to isolate the adipocyte precursor cells from the SVF. Subsequently, this population can be divided into Wt1-expressing and non-expressing fractions, therefore permitting further analysis of the two cell populations, and the mature adipocytes derived from them. In this chapter we outline a method by which adipocyte precursor cells can be isolated, and how, using a specific mouse model, Wt1-expressing and non-expressing cells can be separated.
authors.affiliation: MRC Human Genetics Unit, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, EH4 2XU, UK.
authors.forename: Louise
authors.initials: L
authors.lastname: Cleal
authors.affiliation: MRC Human Genetics Unit, MRC Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh, EH4 2XU, UK. You-Ying.Chau@ed.ac.uk.|British Heart Foundation Centre for Cardiovascular Science, The Queen's Medical Research Institute, University of Edinburgh, 47 Little France Crescent, Edinburgh, EH16 4TJ, UK. You-Ying.Chau@ed.ac.uk.
authors.forename: You-Ying
authors.initials: YY
authors.lastname: Chau
mesh_terms: D017667:Adipocytes / Q000166:cytology* / Q000378:metabolism; D052436:Adipose Tissue, White / Q000166:cytology; D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D005434:Flow Cytometry / Q000379:methods*; D051379:Mice; D012097:Repressor Proteins / Q000378:metabolism*; D025721:WT1 Proteins
chemical_list: D012097:Repressor Proteins; D025721:WT1 Proteins; C000602896:WT1 protein, mouse
keywords: Adipocyte; Adipocyte precursor; Adipose tissue; Flow cytometry; Fluorescence-activated cell sorting; Stromal vascular fraction; Visceral