Fig. 2
Total ADSC secretome and the EV fraction affect three hallmarks of regeneration in vitro. a The impact of ADSC secretome on cell proliferation was assessed using total cell number determination revealing a significant increase in proliferation after exposure of C2C12 cells to the whole secretome. b, c Immunocytochemical analysis demonstrated an increase of C2C12 cell differentiation towards myotubes after whole secretome treatment. d, e Wound healing assay was performed to assess the impact of the secretome on migration of A10 muscle cells. No significant changes in migration of A10 cells exposed to the whole secretome were observed. f In order to study the effects of the total secretome and ADSC-derived EVs on cellular senescence, IMR-90 cells were co-exposed to H2O2 and vehicle (PBS), whole secretome, and the EV-fraction followed by staining for β-Galactosidase activity. Analysis of the number of senescent cells revealed that both total ADSC secretome and the EV fraction protect against cellular senescence. g, h Only the EV fraction (arrows) significantly reduces levels of inflammation in the U251 cell model compared to the TNF-α exposed control cells showing high level of nuclear p65 (arrowheads). Scale bar: 20 μm. p < 0.05 (*), p < 0.01 (**) or p < 0.001 (***). Three separate batches of ADSC secretome and isolated EV fraction were tested