id_1003. DONOR-RELATED VARIABILITY IN EV SECRETION BY WJ-MSCS AND ASCS - CHALLENGES FOR PERSONALIZED CELL-FREE THERAPIES
Monika Rytel1, Sylwia Katarzyna Król2, Anna Sarnowska1
1 Mossakowski Medical Research Institute, Translational Platform for Regenerative Medicine, 5 Pawinskiego St., Warsaw, Poland
2 Mossakowski Medical Research Institute, Department of Neurooncology, 5 Pawinskiego St., Warsaw, Poland
INTRODUCTION: Mesenchymal stem/stromal cells possess immunomodulatory, regenerative, and neuroprotective properties, making them promising candidates for the treatment of neurological disorders. Their therapeutic effects largely depend on their secretome, which includes soluble factors and extracellular vesicles (EVs). EVs are key mediators of intercellular communication, carrying proteins, lipids, nucleic acids, and membrane receptors that can be transferred to recipient cells. Their ability to deliver functional biomolecules has generated strong interest in their use as biomarkers and therapeutic agents.
AIM(S): We hypothesized that MSCs derived from different sources secrete varying numbers of EVs and may therefore differ in their therapeutic potential. Thus, we compared two types of MSCs - WJ-MSCs and ASCs.
METHOD(S): Human umbilical cords (n=5) were obtained from full-term deliveries and MSCs were then mechanically isolated from Wharton’s jelly (WJ-MSCs). Adipose tissue-derived MSCs (ASCs) were isolated enzymatically with collagenase from human adipose tissue (n=5). Next, both WJ-MSCs and ASCs were cultured until 3rd passage, at 5% O2, 5% CO2, 37°C. EVs were then isolated from cell culture medium by ultracentrifugation (300xg for 10 min. 4°C, 2 000xg for 30 min. 4°C, and 100 000xg for 1h 30 min. 4°C) The obtained EVs were suspended in PBS and their number and size was determined by NTA (Nanoparticle Tracking Analysis). EV morphology was analyzed by TEM (Transmission Electron Microscopy).
RESULTS: The analysis revealed that WJ-MSCs and ASCs obtained from different donors secrete different numbers of EVs. EVs isolated from both cell types represented a typical extracellular vesicle-like morphology according to TEM analyses.
CONCLUSIONS: WJ-MSCs and ASCs derived from different donors exhibit donor-dependent variability in the number of secreted EVs, which in the future may potentially have an impact on the efficiency of cell-free therapies dedicated for individual patients.
FINANCIAL SUPPORT: This research was funded by National Science Centre grant number 2022/47/O/NZ3/01739.