Isolation and Fluorescence-Activated Cell Sorting of Murine WT1-Expressing Adipocyte Precursor Cells
- PMID: 27417961
- DOI: 10.1007/978-1-4939-4023-3_7
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.
Keywords: Adipocyte; Adipocyte precursor; Adipose tissue; Flow cytometry; Fluorescence-activated cell sorting; Stromal vascular fraction; Visceral.
Similar articles
-
Flow Cytometry and Lineage Tracing Study for Identification of Adipocyte Precursor Cell (APC) Populations.Methods Mol Biol. 2018;1752:111-121. doi: 10.1007/978-1-4939-7714-7_11. Methods Mol Biol. 2018. PMID: 29564767
-
Flow Cytometric Isolation and Differentiation of Adipogenic Progenitor Cells into Brown and Brite/Beige Adipocytes.Methods Mol Biol. 2017;1566:25-36. doi: 10.1007/978-1-4939-6820-6_4. Methods Mol Biol. 2017. PMID: 28244038
-
Analysis and isolation of adipocytes by flow cytometry.Methods Enzymol. 2014;537:281-96. doi: 10.1016/B978-0-12-411619-1.00015-X. Methods Enzymol. 2014. PMID: 24480352 Free PMC article.
-
Small proliferative adipocytes: identification of proliferative cells expressing adipocyte markers.Endocr J. 2013;60(8):931-9. doi: 10.1507/endocrj.ej13-0141. Epub 2013 Jun 8. Endocr J. 2013. PMID: 23749173 Review.
-
Adipocyte-specific transgenic and knockout models.Methods Enzymol. 2014;537:1-16. doi: 10.1016/B978-0-12-411619-1.00001-X. Methods Enzymol. 2014. PMID: 24480338 Review.
Cited by
-
Identifying Key Genes and Functionally Enriched Pathways of Diverse Adipose Tissue Types in Cattle.Front Genet. 2022 Feb 14;13:790690. doi: 10.3389/fgene.2022.790690. eCollection 2022. Front Genet. 2022. PMID: 35237299 Free PMC article.
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources