Fig. 1

Characterization of MenSC-EV samples by nFC. MenSCs were exposed to different pre-conditioning conditions for 48–72 h. Subsequently, conditioned media from MenSCs were collected in DMEM serum-free media (1%ITS, 1% P/S) for 48 h and EVs were isolated and characterized by combining ultra-filtration and differential centrifugation methods (see details in “Methods” Section). A EV release by MenSCs exposed to different priming conditions. EV release was estimated based on the number of particles released per cell exposed to basal (B, red), pro-inflammatory stimuli (PI, green), physioxia (PHY, blue) or acute hypoxia (AH, yellow) pre-conditioning. Samples from five different donor cell lines were individually analyzed by nano flow-cytometry (nFC). B Median size particles (nm) analyzed in panel A. The line indicates averaged values for n = 5 donors. C Sizing profile of representative pooled samples. A pool of n = 5 donor conditioned media samples was mixed 1:1 and EVs subsequently isolated in order to decrease inter-individual variability. Histograms represent particles detected by nFC using a bin size of 0.5 nm (small EV range, 40–200 nm). Non-linear gaussian fit curves are also plotted in the interest of visualization. EVs, extracellular vesicles; MenSCs, menstrual blood-derived stromal cells; B-EVs, EVs released by basal MenSCs; PI-EVs, EVs released by pro-inflammatory primed MenSCs; PHY-EVs, EVs released by physioxia cultured MenSCs; AH-EVs, EVs released by acute hypoxia cultured MenSCs