FIG. 2.

Young and aged ASCs display similar proliferation, differentiation, and matrix gene expression (A–C) (A) Young and aged ASC proliferation cultured in basal and adipogenic medium was assessed at days 1, 3, 7, and 10 using the CCK-8. Young and aged proliferation was comparable in both basal and adipogenic conditions. (B) Young and aged ASCs exhibit comparable differentiation capacity and morphology in two-dimensional culture. Young and aged ASCs were maintained in CM for 7 days, stained with Alexa Fluor 488 Phalloidin, and counterstained with DAPI. Adipogenic-differentiated young and aged ASCs were maintained for 14 days in adipogenic differentiation medium. Oil Red-O was used to stain for lipid rich vacuoles. (C) Young and aged ASCs exhibit comparable matrix gene expression. Young and aged ASCs were cultured in basal medium and collected at 80% confluency for qRT-PCR analysis. The gene expression of extracellular matrix elements of young ASCs was normalized to aged ASC gene expression. N = 3 biological replicates (all assays), N = 3 technical replicates (proliferation), N = 2 technical replicates (differentiation and qRT-PCR), error bars = SEM (D) TCGA data of young ER-α⁺ breast tumors exhibit significantly elevated matrix gene expression (*P < 0.05) compared to aged. ASC, adipose-derived stromal/stem cell; CCK-8, Cell Counting Kit-8; COL1A1, collagen type I alpha 1; COL3A1, collagen type III alpha 1; COL4A2, collagen type IV alpha 2; COL6A1, collagen type VI alpha 1; FN1, fibronectin-1; LAMA1, laminin subunit alpha 1; LAMA3, laminin subunit alpha 3; LAMB3, laminin subunit beta 3; qRT-PCR, quantitative real time polymerase chain reaction; SEM, standard error of the mean.