AVENTACELL GLOBAL SHOWCASE PRESENTATION Exosome-depleted hPL Supplements Can Support Robust Cell-derived EV Production
Friday, June 2, 2023 12:45 PM - 01:00 PM
Global Showcase Theater
Global Showcase Presentation
Exosome-depleted hPL Supplements Can Support Robust Cell-derived EV Production
Presenters
Claudia Lobato da Silva, PhD, Associate Professor, Institute of Bioengineering & Bioscienc, Stem Cell Engineering Research Group, U. of Lisboa
Cristiana Ulpiano, Instituto Superior Técnico, Universidade de Lisboa
Cell-derived extracellular vesicles (EVs) play an important role in cell-to-cell communication and are emerging as a new platform for promising candidates for both regenerative medicine therapeutics and drug delivery systems based on their unique biological properties. Production process development for scaling-up EV manufacturing will be pivotal to ultimate commercial success. However, exosomes, one of the EV subpopulations, comprised of various growth factors, cytokines, RNAs, and DNAs, are present in some potent cell culture supplements; e.g. fetal bovine serum or human platelet lysate (hPL). These exosomes can modulate mesenchymal stromal cells (MSC) physiology which subsequently affects the quantity and profile of EVs expressed by the MSC and in addition, these supplement EVs can also confound downstream isolation, analysis and collection of the targeted cell-derived EVs; leading to misinterpretation of safety, activity and potency data in EV product R&D. Therefore, qualified ancillary materials for a controlled cell culture environment are crucial for researchers to achieve consistent, batch-to-batch target cell EV expression profiles for GMP production for clinical development and commercialization.
Here we see that Exosome-Depleted UltraGRO™-PURE GI (“ED UG-P GI”, AventaCell Biomedical) the first commercial exosome-depleted hPL cell culture supplement with PRT features, is ideally suited for target cell-derived EV production. >98% of the nanoparticles are removed from the hPL, resulting in minimal hPL nanoparticle contamination for cell-derived EV production (< 0.1% hPL EV presence when applying 5% culture medium). Human MSC maintained in 5% ED UG-P GI supplemented culture medium presented higher cell viability (over 90%), cell numbers and particle concentration for several EV production cycles, compared to cells cultured in serum-free medium, which demonstrated minimal long-term EV production capacity. After switching the cells to 5% ED UG-P GI culture medium, up to 6X1011 MSC-EVs can be harvested in a week using a 175-cm2 culture flask, potentially translating as more than 50 effective doses (ex. 1010/dose) for clinical applications. Moreover, ED UG-P GI supplementation allowed the production of multiple batches of MSC-EVs under stirred conditions in easy scalable spinner flasks. Importantly, isolated MSC-EVs showed the typical size distribution profiles (Nanoparticle Tracking Analysis (NTA) measurements and Transmission Electron Microscopy (TEM) imaging) and expressed EV-positive protein markers (CD63, CD9 and HSP70). In addition, expanded MSC maintained the characteristic immunophenotypic profile at the end of the culture period.
These iBB research results conclude that exosome-depleted hPL (ED UG-P GI) is feasible for MSC-EV product R&D to prolong cell viability for robust therapeutic EV production process development. Therefore, exosome-depleted hPL (ED UG-P GI) is a promising supplement which can be utilized in EV research and development, for disease process, cancer product development, tissue regeneration as well as the scaled-up GMP cell-derived EV manufacturing for clinical and commercial applications.
Presenters
Claudia Lobato da Silva, PhD, Associate Professor, Institute of Bioengineering & Bioscienc, Stem Cell Engineering Research Group, U. of Lisboa
Cristiana Ulpiano, Instituto Superior Técnico, Universidade de Lisboa
Cell-derived extracellular vesicles (EVs) play an important role in cell-to-cell communication and are emerging as a new platform for promising candidates for both regenerative medicine therapeutics and drug delivery systems based on their unique biological properties. Production process development for scaling-up EV manufacturing will be pivotal to ultimate commercial success. However, exosomes, one of the EV subpopulations, comprised of various growth factors, cytokines, RNAs, and DNAs, are present in some potent cell culture supplements; e.g. fetal bovine serum or human platelet lysate (hPL). These exosomes can modulate mesenchymal stromal cells (MSC) physiology which subsequently affects the quantity and profile of EVs expressed by the MSC and in addition, these supplement EVs can also confound downstream isolation, analysis and collection of the targeted cell-derived EVs; leading to misinterpretation of safety, activity and potency data in EV product R&D. Therefore, qualified ancillary materials for a controlled cell culture environment are crucial for researchers to achieve consistent, batch-to-batch target cell EV expression profiles for GMP production for clinical development and commercialization.
Here we see that Exosome-Depleted UltraGRO™-PURE GI (“ED UG-P GI”, AventaCell Biomedical) the first commercial exosome-depleted hPL cell culture supplement with PRT features, is ideally suited for target cell-derived EV production. >98% of the nanoparticles are removed from the hPL, resulting in minimal hPL nanoparticle contamination for cell-derived EV production (< 0.1% hPL EV presence when applying 5% culture medium). Human MSC maintained in 5% ED UG-P GI supplemented culture medium presented higher cell viability (over 90%), cell numbers and particle concentration for several EV production cycles, compared to cells cultured in serum-free medium, which demonstrated minimal long-term EV production capacity. After switching the cells to 5% ED UG-P GI culture medium, up to 6X1011 MSC-EVs can be harvested in a week using a 175-cm2 culture flask, potentially translating as more than 50 effective doses (ex. 1010/dose) for clinical applications. Moreover, ED UG-P GI supplementation allowed the production of multiple batches of MSC-EVs under stirred conditions in easy scalable spinner flasks. Importantly, isolated MSC-EVs showed the typical size distribution profiles (Nanoparticle Tracking Analysis (NTA) measurements and Transmission Electron Microscopy (TEM) imaging) and expressed EV-positive protein markers (CD63, CD9 and HSP70). In addition, expanded MSC maintained the characteristic immunophenotypic profile at the end of the culture period.
These iBB research results conclude that exosome-depleted hPL (ED UG-P GI) is feasible for MSC-EV product R&D to prolong cell viability for robust therapeutic EV production process development. Therefore, exosome-depleted hPL (ED UG-P GI) is a promising supplement which can be utilized in EV research and development, for disease process, cancer product development, tissue regeneration as well as the scaled-up GMP cell-derived EV manufacturing for clinical and commercial applications.
Claudia Lobato da Silva PhD
Associate Professor
Associação do Instituto Superior Técnico para a investigação e Desenvolvimento (IST-ID)
Presenter
Associate Professor
Associação do Instituto Superior Técnico para a investigação e Desenvolvimento (IST-ID)
Presenter
