Hollow-fiber bioreactor production of extracellular vesicles from human bone marrow mesenchymal stromal cells yields nanovesicles that mirrors the immuno-modulatory antigenic signature of the producer cell.
Extracellular vesicles
Glycan
Hollow-fiber bioreactor system
Human mesenchymal stromal cells
Immune-profiling
cGMP-compliant environment
Journal
Stem cell research & therapy
ISSN: 1757-6512
Titre abrégé: Stem Cell Res Ther
Pays: England
ID NLM: 101527581
Informations de publication
Date de publication:
12 02 2021
12 02 2021
Historique:
received:
25
08
2020
accepted:
25
01
2021
entrez:
13
2
2021
pubmed:
14
2
2021
medline:
9
7
2021
Statut:
epublish
Résumé
Extracellular vesicles (EVs) produced by human bone marrow-derived mesenchymal stromal cells (hBM-MSCs) are currently investigated for their clinical effectiveness towards immune-mediated diseases. The large amounts of stem cell-derived EVs required for clinical testing suggest that bioreactor production systems may be a more amenable alternative than conventional EV production methods for manufacturing products for therapeutic use in humans. To characterize the potential utility of these systems, EVs from four hBM-MSC donors were produced independently using a hollow-fiber bioreactor system under a cGMP-compliant procedure. EVs were harvested and characterized for size, concentration, immunophenotype, and glycan profile at three separate intervals throughout a 25-day period. Bioreactor-inoculated hBM-MSCs maintained high viability and retained their trilineage mesoderm differentiation capability while still expressing MSC-associated markers upon retrieval. EVs collected from the four hBM-MSC donors showed consistency in size and concentration in addition to presenting a consistent surface glycan profile. EV surface immunophenotypic analyses revealed a consistent low immunogenicity profile in addition to the presence of immuno-regulatory CD40 antigen. EV cargo analysis for biomarkers of immune regulation showed a high abundance of immuno-regulatory and angiogenic factors VEGF-A and IL-8. Significantly, EVs from hBM-MSCs with immuno-regulatory constituents were generated in a large-scale system over a long production period and could be frequently harvested with the same quality and quantity, which will circumvent the challenge for clinical application.
Sections du résumé
BACKGROUND
Extracellular vesicles (EVs) produced by human bone marrow-derived mesenchymal stromal cells (hBM-MSCs) are currently investigated for their clinical effectiveness towards immune-mediated diseases. The large amounts of stem cell-derived EVs required for clinical testing suggest that bioreactor production systems may be a more amenable alternative than conventional EV production methods for manufacturing products for therapeutic use in humans.
METHODS
To characterize the potential utility of these systems, EVs from four hBM-MSC donors were produced independently using a hollow-fiber bioreactor system under a cGMP-compliant procedure. EVs were harvested and characterized for size, concentration, immunophenotype, and glycan profile at three separate intervals throughout a 25-day period.
RESULTS
Bioreactor-inoculated hBM-MSCs maintained high viability and retained their trilineage mesoderm differentiation capability while still expressing MSC-associated markers upon retrieval. EVs collected from the four hBM-MSC donors showed consistency in size and concentration in addition to presenting a consistent surface glycan profile. EV surface immunophenotypic analyses revealed a consistent low immunogenicity profile in addition to the presence of immuno-regulatory CD40 antigen. EV cargo analysis for biomarkers of immune regulation showed a high abundance of immuno-regulatory and angiogenic factors VEGF-A and IL-8.
CONCLUSIONS
Significantly, EVs from hBM-MSCs with immuno-regulatory constituents were generated in a large-scale system over a long production period and could be frequently harvested with the same quality and quantity, which will circumvent the challenge for clinical application.
Identifiants
pubmed: 33579358
doi: 10.1186/s13287-021-02190-3
pii: 10.1186/s13287-021-02190-3
pmc: PMC7880218
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
127Références
J Extracell Vesicles. 2018 Feb 28;7(1):1442088
pubmed: 29535850
Exp Mol Med. 2019 Mar 15;51(3):1-12
pubmed: 30872574
Stem Cells. 2014 Jun;32(6):1380-9
pubmed: 24497003
Stem Cells. 2017 Apr;35(4):851-858
pubmed: 28294454
Cytotherapy. 2012 Nov;14(10):1159-63
pubmed: 23066784
Cell Prolif. 2019 Jul;52(4):e12604
pubmed: 31069891
Blood. 1991 Dec 1;78(11):2848-53
pubmed: 1720038
J Extracell Vesicles. 2014 Dec 22;3:25465
pubmed: 25536933
J Extracell Vesicles. 2015 May 14;4:27066
pubmed: 25979354
J Extracell Vesicles. 2018 Nov 23;7(1):1535750
pubmed: 30637094
Biomark Res. 2019 Mar 18;7:6
pubmed: 30923617
Cell Stem Cell. 2018 Jun 01;22(6):824-833
pubmed: 29859173
Drug Discov Today. 2016 May;21(5):740-65
pubmed: 26821133
Blood. 1991 Jul 1;78(1):55-62
pubmed: 2070060
Blood. 1995 May 1;85(9):2422-35
pubmed: 7537114
Nat Rev Drug Discov. 2013 May;12(5):347-57
pubmed: 23584393
Front Cell Dev Biol. 2016 Aug 22;4:83
pubmed: 27597941
Cell Biosci. 2019 Feb 15;9:19
pubmed: 30815248
Cytotherapy. 2013 Nov;15(11):1323-39
pubmed: 23992670
Biochem Biophys Res Commun. 2013 Jan 4;430(1):325-30
pubmed: 23146633
Stem Cell Res Ther. 2017 Nov 2;8(1):247
pubmed: 29096714
Cytotherapy. 2014 Aug;16(8):1048-58
pubmed: 24726657
Exp Hematol. 2002 Jul;30(7):783-91
pubmed: 12135677
Proc Natl Acad Sci U S A. 2010 Aug 3;107(31):13724-9
pubmed: 20643923
Stem Cells. 2018 Mar;36(3):434-445
pubmed: 29239062
Biomaterials. 2016 Oct;105:195-205
pubmed: 27522254
Stem Cells Int. 2018 Jul 31;2018:7357213
pubmed: 30154865
Blood. 1990 Jun 15;75(12):2417-26
pubmed: 1693532
Biochem Biophys Res Commun. 2009 Apr 17;381(4):676-81
pubmed: 19250924
Biomaterials. 2017 Aug;135:62-73
pubmed: 28494264
Leukemia. 2014 Apr;28(4):970-3
pubmed: 24445866
Cell Transplant. 2013;22(11):1981-2000
pubmed: 23107560
Stem Cells Int. 2013;2013:698076
pubmed: 24194767
Stem Cells Dev. 2016 Nov 15;25(22):1755-1766
pubmed: 27539404
J Extracell Vesicles. 2015 Dec 31;4:30087
pubmed: 26725829
Stem Cells Dev. 2020 Jun 15;29(12):747-754
pubmed: 32380908
Stem Cell Res Ther. 2019 Dec 18;10(1):401
pubmed: 31852509
Stem Cells Int. 2018 Dec 24;2018:1310904
pubmed: 30675166
Cytotherapy. 2017 May;19(5):629-639
pubmed: 28366194
Cell Tissue Res. 2020 Apr;380(1):93-105
pubmed: 31889209
Biotechnol Lett. 2019 Jan;41(1):1-25
pubmed: 30368691
Stem Cell Res Ther. 2019 Feb 7;10(1):51
pubmed: 30732645
J Clin Invest. 2016 Apr 1;126(4):1139-43
pubmed: 27035805
Cytotherapy. 2006;8(4):315-7
pubmed: 16923606
Sci Rep. 2019 Mar 7;9(1):3771
pubmed: 30846806
Nat Biotechnol. 2014 Mar;32(3):252-60
pubmed: 24561556
J Nanobiotechnology. 2018 Oct 16;16(1):81
pubmed: 30326899
Methods Mol Biol. 2016;1416:389-412
pubmed: 27236685
Proc Natl Acad Sci U S A. 2016 Feb 23;113(8):E968-77
pubmed: 26858453
Oncogene. 2019 Jun;38(26):5158-5173
pubmed: 30872795
Nat Cell Biol. 2019 Jan;21(1):9-17
pubmed: 30602770
J Extracell Vesicles. 2018 Sep 26;7(1):1522236
pubmed: 30275938
Biochimie. 2013 Dec;95(12):2212-21
pubmed: 23810910
Adv Mater. 2019 Feb;31(7):e1806214
pubmed: 30589121