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Scalable production of human mesenchymal stromal cell-derived extracellular vesicles under serum-/xeno-free conditions in a microcarrier-based bioreactor culture system

2020Journal article Open access
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dc.contributor.authorFuzeta, Miguel de Almeida
dc.contributor.authorBernardes, Nuno
dc.contributor.authorOliveira, Filipa D.
dc.contributor.authorCosta, Ana Catarina
dc.contributor.authorFernandes-Platzgummer, Ana
dc.contributor.authorFarinha, José Paulo
dc.contributor.authorRodrigues, Carlos A.V.
dc.contributor.authorJung, Sunghoon
dc.contributor.authorTseng, Rong-Jeng
dc.contributor.authorMilligan, William
dc.contributor.authorLee, Brian
dc.contributor.authorCastanho, Miguel A. R. B.
dc.contributor.authorGaspar, Diana
dc.contributor.authorCabral, Joaquim M.S.
dc.contributor.authorda Silva, Cláudia Lobato
dc.date.accessioned2021-04-08T11:28:02Z
dc.date.available2021-04-08T11:28:02Z
dc.date.issued2020
dc.descriptionCopyright © 2020 de Almeida Fuzeta, Bernardes, Oliveira, Costa, Fernandes-Platzgummer, Farinha, Rodrigues, Jung, Tseng, Milligan, Lee, Castanho, Gaspar, Cabral and da Silva. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.pt_PT
dc.description.abstractMesenchymal stromal cells (MSC) hold great promise for tissue engineering and cell-based therapies due to their multilineage differentiation potential and intrinsic immunomodulatory and trophic activities. Over the past years, increasing evidence has proposed extracellular vesicles (EVs) as mediators of many of the MSC-associated therapeutic features. EVs have emerged as mediators of intercellular communication, being associated with multiple physiological processes, but also in the pathogenesis of several diseases. EVs are derived from cell membranes, allowing high biocompatibility to target cells, while their small size makes them ideal candidates to cross biological barriers. Despite the promising potential of EVs for therapeutic applications, robust manufacturing processes that would increase the consistency and scalability of EV production are still lacking. In this work, EVs were produced by MSC isolated from different human tissue sources [bone marrow (BM), adipose tissue (AT), and umbilical cord matrix (UCM)]. A serum-/xeno-free microcarrier-based culture system was implemented in a Vertical-WheelTM bioreactor (VWBR), employing a human platelet lysate culture supplement (UltraGROTM-PURE), toward the scalable production of MSC-derived EVs (MSC-EVs). The morphology and structure of the manufactured EVs were assessed by atomic force microscopy, while EV protein markers were successfully identified in EVs by Western blot, and EV surface charge was maintained relatively constant (between −15.5 ± 1.6 mV and −19.4 ± 1.4 mV), as determined by zeta potential measurements. When compared to traditional culture systems under static conditions (T-flasks), the VWBR system allowed the production of EVs at higher concentration (i.e., EV concentration in the conditioned medium) (5.7-fold increase overall) and productivity (i.e., amount of EVs generated per cell) (3-fold increase overall). BM, AT and UCM MSC cultured in the VWBR system yielded an average of 2.8 ± 0.1 × 1011, 3.1 ± 1.3 × 1011, and 4.1 ± 1.7 × 1011 EV particles (n = 3), respectively, in a 60 mL final volume. This bioreactor system also allowed to obtain a more robust MSC-EV production, regarding their purity, compared to static culture. Overall, we demonstrate that this scalable culture system can robustly manufacture EVs from MSC derived from different tissue sources, toward the development of novel therapeutic products.pt_PT
dc.description.sponsorshipunding received by iBB-Institute for Bioengineering and Biosciences from the Portuguese Foundation for Science and Technology (FCT) (UID/BIO/04565/2020) and through the projects PTDC/EQU-EQU/31651/2017, PTDC/BBB-BQB/1693/2014, and PTDC/BTM-SAL/31057/2017 is acknowledged. Funding received from POR de Lisboa 2020 through the project PRECISE – Accelerating progress toward the new era of precision medicine (Project N. 16394) is also acknowledged. MAF (PD/BD/128328/2017) and FO (PD/BD/135046/2017) acknowledge FCT for the Ph.D. fellowships and DG (SFRH/BPD/109010/2015) for the Post-Doctoral fellowship.pt_PT
dc.description.versioninfo:eu-repo/semantics/publishedVersionpt_PT
dc.identifier.citationFront Cell Dev Biol. 2020 Nov 3;8:553444.pt_PT
dc.identifier.doi10.3389/fcell.2020.553444pt_PT
dc.identifier.eissn2296-634X
dc.identifier.urihttp://hdl.handle.net/10451/47284
dc.language.isoengpt_PT
dc.peerreviewedyespt_PT
dc.publisherFrontierspt_PT
dc.relationUID/BIO/04565/2020pt_PT
dc.relationCell-cell communication in metastatic breast cancer: a control point that can lead to breast cancer treatment
dc.relation16394pt_PT
dc.relationDoctoral Program MIT Portugal Bio-Engineering Systems Exosomes from Human Mesenchymal Stem/stromal cells for the Delivery of Anticancer Peptides to treat breast cancer
dc.relationExosomes as drug delivery systems in metastatic breast cancer
dc.relationCell-cell direct/indirect communication in metastatic breast cancer as a control point in oncotherapy
dc.relation.publisherversionhttps://www.frontiersin.org/journals/cell-and-developmental-biologypt_PT
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/pt_PT
dc.subjectExtracellular vesiclespt_PT
dc.subjectMesenchymal stromal cells (MSC)pt_PT
dc.subjectScalable productionpt_PT
dc.subjectBioreactorspt_PT
dc.subjectSerum- /xenogeneic-freept_PT
dc.titleScalable production of human mesenchymal stromal cell-derived extracellular vesicles under serum-/xeno-free conditions in a microcarrier-based bioreactor culture systempt_PT
dc.typejournal article
dspace.entity.typePublication
oaire.awardTitleCell-cell communication in metastatic breast cancer: a control point that can lead to breast cancer treatment
oaire.awardTitleDoctoral Program MIT Portugal Bio-Engineering Systems Exosomes from Human Mesenchymal Stem/stromal cells for the Delivery of Anticancer Peptides to treat breast cancer
oaire.awardTitleExosomes as drug delivery systems in metastatic breast cancer
oaire.awardTitleCell-cell direct/indirect communication in metastatic breast cancer as a control point in oncotherapy
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FEQU-EQU%2F31651%2F2017/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/3599-PPCDT/PTDC%2FBBB-BQB%2F1693%2F2014/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/9471 - RIDTI/PTDC%2FBTM-SAL%2F31057%2F2017/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//PD%2FBD%2F128328%2F2017/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//PD%2FBD%2F135046%2F2017/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//SFRH%2FBPD%2F109010%2F2015/PT
oaire.citation.titleFrontiers in Cell and Developmental Biologypt_PT
oaire.citation.volume8pt_PT
oaire.fundingStream3599-PPCDT
oaire.fundingStream3599-PPCDT
oaire.fundingStream9471 - RIDTI
person.familyNamede Almeida Fuzeta
person.familyNameBernardes
person.familyNameFernandes-Platzgummer
person.familyNameVitorino Rodrigues
person.familyNameCastanho
person.familyNameGaspar
person.familyNameCabral
person.givenNameMiguel
person.givenNameNuno
person.givenNameAna
person.givenNameCarlos André
person.givenNameMiguel
person.givenNameDiana
person.givenNameJoaquim M.S.
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person.identifier.ridF-7826-2014
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person.identifier.scopus-author-id38662250700
person.identifier.scopus-author-id56605575600
person.identifier.scopus-author-id55180303000
person.identifier.scopus-author-id7201350203
project.funder.identifierhttp://doi.org/10.13039/501100001871
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rcaap.rightsopenAccesspt_PT
rcaap.typearticlept_PT
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