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. 2014:537:31-46.
doi: 10.1016/B978-0-12-411619-1.00003-3.

Isolation and study of adipocyte precursors

Christopher D Church  1 , Ryan Berry  2 , Matthew S Rodeheffer  3
Affiliations

Isolation and study of adipocyte precursors

Christopher D Church et al. Methods Enzymol. 2014.

Abstract

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.

Keywords: Adipocyte precursor; Adipogenesis; Adipose tissue; Preadipocytes; Progenitor; Stem cell; Stromal vascular.

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Figures

Figure 1
Figure 1
Separation of SVF from Adipocytes
Figure 2
Figure 2. Flow Cytometry Gating of Adipocyte Precursor Cells
Each plot displays only the cell population from the populations gated in the leftward plot, which is also indicated at the top of each plot.
Figure 3
Figure 3. Adipocyte Precursor Cell Purity with Minimal Cell Surface Markers
Data (n=4) are expressed as mean±SEM.
Figure 4
Figure 4. Differentiation of Primary Adipocyte Precursor Cells
A. Day 7 Subcutaneous WAT (SWAT), B. Day 7 Epigonadal WAT (EWAT), C. Day 7 Oil Red O quantification. INS, insulin only differentiation. MDI (adipogenic cocktail containing IBMX, dexamethasone and insulin).
Figure 5
Figure 5. Isolation of Adipocytes
A. Intact adipocytes with single DAPI positive nuclei within a cell membrane B. Lipid ghosts without DAPI positive nuclei. White scale bar represents 100 μm.
See this image and copyright information in PMC

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