Metabolic Phenotyping of Adipose-Derived Stem Cells Reveals a Unique Signature and Intrinsic Differences between Fat Pads

Figure 5

Quantitative comparison of S- and V-ASC secretomes: S- and V-ASC at 75% confluency were cultivated for 72 h in fresh culture medium containing pyruvate to reach confluency. Cells were counted as supernatants were collected for NMR analysis. Variations of the concentrations after deduction of the medium values are represented without normalization to the cell number. (a) Metabolites representing glycolysis (left panel) and other metabolites (right panel) showing significant differences between S- and V-ASC (). (b) Number of cells obtained after 72 h of culture with pyruvate. Cells were counted after collection of the culture supernatants (). (c) Cumulative growth curves: S- and V-ASC were cultivated in complete culture medium (see Materials and Methods) from the isolation to the 4^th passage. The number of cells after each passage is represented (). (d) Comparison of Ucp2 mRNA expression in S- and V-ASC. Ucp2 mRNA levels were quantified by RT-qPCR in S- and V-ASC harvested at the second passage. Values are normalized on the expression of the housekeeping gene RS17 (). (e) Comparison of UCP2 protein content in S- and V-ASC cell lysates by western blotting. Cell lysates were from S- and V-ASC harvested at the second passage. Images (top) were obtained with the ChemiDoc XRS+ imaging system (Bio-Rad) and were analyzed (bottom) with the Quantity One software (Bio-Rad). Values were normalized relatively to the α-tubulin protein expression (). All results are ; statistics are from -tests: , , and .

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