Determining stemness and exosome production from nutrient-dependent cells: Influence of the molecular weight cut-off in hollow fiber membrane systems.

This study aimed to optimize parameters for cultivating adipose-derived stem cells (ADSCs) in hollow fiber membrane systems. Enhanced ADSC cultivation was achieved in this study by utilizing a higher cell seeding rate (10 mL/min) and fibronectin coating (8 μg/cm²) on the membrane. A pulsatile flow perfusion (30 min on / 3 h off) for 24 h, followed by continuous flow perfusion at a lower rate (<20 mL/min), was employed. Notably, a two-day medium change maintained CD73 expression (79.26 %) compared to scenarios without changes (1.92 %) over eight days. Using a membrane cut-off of 20 kDa and <0.1 μm, a 1.4- and 2.8-fold increase in proliferated cells was observed; however, CD73 expression decreased by 10.07 and 33.63 %, respectively. The membrane with a pore size of <0.1 μm exhibited 1.4-fold higher exosome particle numbers. In conclusion, nutrient supplementation emerged as crucial for optimal ADSC cultivation in hollow fiber systems. [Display omitted] • This study aimed to optimize parameters for cultivating adipose-derived stem cells (ADSCs) in hollow fiber membrane systems. • Investigated hollow fiber feasibility for stem cell cultivation and exosome production. • Two-day medium change increased cell proliferation by 3 times. • Stemness marker increases 3x on <0.1 µm MWCO fiber membrane vs. 20 kDa membrane. • Proved nutrient supply significantly influences stemness, proliferation, and exosome production. [ABSTRACT FROM AUTHOR]