Forschung & Innovation
Publikationen
Heparin Can Augment Extracellular Vesicle Yield by Inhibiting Reuptake in Donor Cells During Cell Propagation
PMU Autor*innen
Andre Cronemberger Andrade, Rodolphe Poupardin, Martin Wolf, Liliia Paniushkina, Nicole Maeding, Heide-Marie Binder, Janine Zwicklhuber, Sarah Hochmann, Katharina Schallmoser, Dirk Strunk
Alle Autor*innen
Andre Cronemberger Andrade, Rodolphe Poupardin, Martin Wolf, Liliia Paniushkina, Karim Selmi, Deepika Kundully Suresh, Nicole Maeding, Heide-Marie Binder, Janine Zwicklhuber, Sarah Hochmann, Andre Gorgens, Samir El Andaloussi, Katharina Schallmoser, Dirk Strunk
Fachzeitschrift
Journal of Extracellular Vesicles
Kurzfassung
Extracellular vesicles (EVs) mediate cell-to-cell communication in development, health and disease. Heparin is a common component in human induced pluripotent stem cell (hiPSC) media but its impact on hiPSC-EVs beyond stabilizing growth factors is unknown. Published evidence indicates heparin dose-dependent EV uptake inhibition in heterotypic target cells. We asked if heparin affects hiPSC-EV yield, storage, phenotype, or function. We used large-scale hiPSC-EV propagation to obtain pro-angiogenic hiPSC-EVs in presence or absence of escalating heparin doses and determined EV quantity, identity and functionality. Heparin delayed hiPSC colony-confluence without significantly affecting hiPSC viability, pluripotency markers and proliferation. Significantly elevated tetraspanin(+)/flotillin(+) hiPSC-EV number (mean 367 +/- 31 EVs/cell/24 h at 200 IU heparin/mL) enabled efficient purification of > 10(11)/mL functional EVs at superior particle/protein ratio > 10(10)/mg. Heparin dose-dependently inhibited EV uptake in hiPSC and endothelial progenitors, comparable to 5 IU/mL flavobacterium heparinase, and increased EV recovery after storage in NaCl but not in cell-conditioned media. EVs obtained from hiPSC at 1% O-2 with 200 IU/mL heparin maintained superior angiogenesis similarly to control hiPSC-EVs obtained without heparin. We conclude that heparin inhibits hiPSC-EV reuptake and stabilizes NaCl-cryopreserved EVs thus boosting EV recovery while maintaining pro-angiogenic function. Heparin presence and dose should therefore be documented particularly in PSC media.
Keywords
HEPARIN, Regenerative medicine, EV function, EV uptake, extracellular vesicle (EV), human induced pluripotent stem cells (hiPSC)