Figure 1.
Isolation and characterization of human ASC.
A. Morphology of human ASC populations, as detected under bright field. B. Expression of specific markers measured by qRT-PCR. Cells were harvested at passage 0, and RNA was analyzed for markers of adipose-derived stem cells (CD105, CD44, CD49d), human leukocytes and macrophages (CD45, CD11b), and mature endothelial cells (CD31).+or – signs indicate the relative levels of marker expression. Results are from 5 independent adipose tissue donors.
Figure 2.
mRNA expression levels of contaminating mesenchymal lineage markers (CD45, CD31, CD11b) (A) and adipose-derived stem cell markers (CD105, CD44, CD49d) (B), measured by qRT-PCR.
ASCSVF were analyzed at passage 0 (P0) and at passage 4 (P4). Open bars, Sc-ASC; filled bars, V-ASC. *p<0.001 vs. P0. Results are from 5 independent adipose tissue donors.
Figure 3.
Venn diagrams summarizing the number of differentially expressed genes in Sc and V ASC populations.
A. Genes found to be differentially expressed by comparing Sc-ASC and V-ASC subsets (inter-depot analysis). B. Genes found to be differentially expressed by comparing ASCSVF, ASCBottom, and ASCCeiling from the Sc adipose tissue (Sc intra-depot analysis). C. Genes found to be differentially expressed by comparing ASCSVF, ASCBottom, and ASCCeiling from the V adipose tissue (V intra-depot analysis). In each panel, figures for conjoint (and non-conjoint) differentially expressed genes are also indicated.
Figure 4.
Global views of gene expression utilizing the Principal Component Analysis (PCA).
A. PCA comparing gene expression of Sc-ASC (blue) and V-ASC (red) subsets. B. PCA comparing gene expression of ASCSVF (red), ASCBottom (blue), and ASCCeiling (green) from Sc and V fat depots, respectively. PCA analyses were performed by Partek GS software using default settings that include a threshold to remove low background level intensities. PCA percent mapping on the top of each plot indicates the explained variability on the first coordinate.
Table 1.
Significant top list of associated network functions from IPA in Sc and V ASCs.
Table 2.
Significant top list of canonical pathways from IPA in Sc and V ASCs.
Figure 5.
Quantitative analysis of specific genes previously identified in the microarray analysis and found to be differentially expressed in the distinct ASC subsets.
mRNA levels were analyzed by qRT-PCR, as described under Methods. Open bars, Sc-ASC; filled bars, V-ASC. All data represent mean ± SE from 9 independent adipose tissue donors. *p<0.05 vs. V-ASC; #p<0.05 vs. other ASC populations from the same adipose tissue depot (ANOVA test followed by Fisher’s post-hoc test). SVF, stromo-vascular; B, bottom; C, ceiling.
Figure 6.
Release of cytokines from Sc-ASC and V-ASC populations.
Culture medium from Sc-ASC (open bars) and V-ASC (filled bars) subsets (106 cells) was collected after a 16-h period, and levels of specific cytokines were determined using the multiplex technique, as described under Materials and Methods. Data represent the mean ± SE of results from 9 independent adipose tissue donors. *p<0.05 vs. V; #p<0.05 vs. other ASC subsets from the same adipose tissue depot (ANOVA test followed by Fisher’s post-hoc test).