Fig. 1.
A 7-day culture timeline of hBM-MSCs designed to include a 48-h production time of sEVs. a A 7-day culture timeline of hBM-MSCs seeded at 1.4 × 105 per T-175 flask at day #1 was established. The hBM-MSCs were grown under normal medium condition (α-MEM with 15% MSC-screened FBS) until day 5, where a medium change was included at day 4. At day 5, the culture medium was changed to an exosome-depleted medium (α-MEM with 15% exosome-depleted FBS) to perform a 48-h hBM-MSC-sEV production time. At day 7, the 48-h CCM produced under exosome-depleted FBS medium condition was collected and frozen at − 80 °C until ready for hBM-MSC-sEV isolation (ED: exosome depleted). b Representative phase contrast images at × 100 magnification taken for all five hBM-MSC donors (i–v) on day 7 showed typical MSC fibroblastic appearance (scale bar = 200 μm). Similar hBM-MSC c counts of viable cells (mean ± SD) per flask, d percentage viability (mean ± SD), and e growth percentage (mean ± SD) were measured. The data represent five hBM-MSC donors (n = 5 donors; D1-D5) analyzed in three independent trials (n = 3 experiments), each trial conducted with two technical replicates (n = 2 technical replicates)