Figure 1.
Adipose tissue depot weight and SVF cell counts.
(A) Whole tissue weight. (B) Total number of SVF cells isolated from the digestion of each tissue. (C) Number of SVF cells isolated from the digestion of each milligram (mg) of adipose tissue. pmWAT; parametrial WAT, rpWAT; retroperitoneal WAT, omWAT; omental WAT. a,b,c Unlike letters indicate significance, p<0.05.
Figure 2.
Flow cytometric analysis gating strategy.
(A) Digested tissue samples were subjected to PI live/dead cell exclusion and CD45+ leukocytes were divided into R1 (lymphocytes) and R2 (mono-, granulocytes) gates based on forward/side scatter followed by doublet exclusion. (B) CD3+ T cells from R1 were further subclassified either as CD4+ TH, CD8+ TC or DN (double negative). (C) CD19+ B cells from R1 and R2 were further subclassified into CD11b+ B1 or CD11b− B2. (D) Monocytic populations from R2 were classified based on CD11b, F4/80 and CD11c (not shown) staining, and further subdivided based on activation markers.
Figure 3.
Flow cytometric analysis of leukocyte populations in the SVF of (A) pmWAT, (B) rpWAT, (C) omWAT and (D) PSF.
Natural killer (NK) cells include mNKs and preNKs, dendritic cells (DC) include DC (I) from R1 and DC (II) from R2, macrophages include large (LPMs) and small (SPM) peritoneal macrophages, monocytes include monocytes (I) (CD11b+F4/80−) and monocytes (II) (CD11bloF4/80−) populations, and “other” includes regulatory T cells (Treg), CD3+CD4−CD8−NK1.1− subset, and myeloid precursors from R1, as well as neutrophils (PMNs), and pre-B or –macrophage cells (PreBoMs) from R2.
Table 1.
Leukocyte Characterization of Immune Microenvironments.
Figure 4.
Myeloid populations found in pmWAT, rpWAT, omWAT and PSF.
(A) Resident myeloid populations differ in a microenvironment-specific manner. (B) Differential CD11b and F4/80 expression displayed as a percentage of total R2 gate. (C) Activation status of respective myeloid populations differ in a microenvironment-specific manner.
Figure 5.
Identification of a unique CD45+ population in the PSF.
Pre-B or –macrophage cells (PreBoMs) found within the R2 gate express CD19+CD11bhiF480+CD93+CD69+CD1d+CD5+ phenotype. This subset was not found in WAT.
Figure 6.
Flow cytometric analysis of stem and progenitor cell markers.
(A) Percentage of CD45− cells that are positive for indicated markers. (B) Total number of SVF cells positive for selected markers in pmWAT and omWAT.
Figure 7.
Heat map of mRNA expression profiles of adipose tissue depots (based on dCT values).
Table 2.
Gene Expression Profiles of Immune Microenvironments.