CM from intact hAM: an easily obtained product with relevant implications for translation in regenerative medicine.
Amniotic membrane
Conditioned medium
Immune functional assays
Immunomodulation
Lyophilization
Secretome
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 10 2021
12 10 2021
Historique:
received:
08
04
2021
accepted:
23
09
2021
entrez:
13
10
2021
pubmed:
14
10
2021
medline:
30
10
2021
Statut:
epublish
Résumé
It is now well established that factors (free or in extracellular vesicles) secreted by mesenchymal stromal cells (MSC) are important mediators of MSC regenerative actions. Herein we produced the secretome (conditioned medium, CM) from MSC isolated from the amniotic membrane (hAMSC) and CM from the intact amniotic membrane (hAM, no manipulation or enzymatic digestion) in order to potentially identify an effective, easy and less expensive secretome to produce for potential applications in regenerative medicine. Given that immunomodulation is a key mechanism of action through which hAMSC contributes to tissue regeneration, we used a comprehensive panel of in vitro immunomodulatory tests to compare the CMs. Amniotic membranes were either cut into fragments or used for hAMSC isolation. CMs from hAMSC at passages 0 and 2 were collected after a standard 5-day culture while CM from hAM was collected after a 2- and 5-day culture. Immunomodulation was assessed in terms of PBMC and T-cell proliferation, T-cell subset polarization, T-regulatory cell induction, cell cytotoxicity and monocyte differentiation toward antigen-presenting cells. Furthermore, we performed a comparison between CM obtained from single donors and pooled CM. We also assessed the impact of lyophilization on the immunomodulatory properties of CM. We demonstrate that CM from hAM has comparable immunomodulatory properties to CM from hAMSC at passages 0 and 2. Furthermore, we demonstrate that pooled CMs have similar effects when compared to CM from single donors used separately. Finally, we demonstrate that lyophilization does not alter the in vitro immunomodulatory properties of CM from hAM and hAMSC. The results presented herein support the possibility to produce secretome from intact hAM and open the prospect to highly improve the scalability of the GMP production process while reducing the costs and time related to the process of cell isolation and expansion. Moreover, the possibility of having a lyophilized secretome that maintains its original properties would allow for a ready-to-use product with easier handling, shipping and storage. The use of a lyophilized product will also facilitate clinicians by permitting customized reconstitution volumes and methods according to the most suitable formula required by the clinical application.
Sections du résumé
BACKGROUND
It is now well established that factors (free or in extracellular vesicles) secreted by mesenchymal stromal cells (MSC) are important mediators of MSC regenerative actions. Herein we produced the secretome (conditioned medium, CM) from MSC isolated from the amniotic membrane (hAMSC) and CM from the intact amniotic membrane (hAM, no manipulation or enzymatic digestion) in order to potentially identify an effective, easy and less expensive secretome to produce for potential applications in regenerative medicine. Given that immunomodulation is a key mechanism of action through which hAMSC contributes to tissue regeneration, we used a comprehensive panel of in vitro immunomodulatory tests to compare the CMs.
METHODS
Amniotic membranes were either cut into fragments or used for hAMSC isolation. CMs from hAMSC at passages 0 and 2 were collected after a standard 5-day culture while CM from hAM was collected after a 2- and 5-day culture. Immunomodulation was assessed in terms of PBMC and T-cell proliferation, T-cell subset polarization, T-regulatory cell induction, cell cytotoxicity and monocyte differentiation toward antigen-presenting cells. Furthermore, we performed a comparison between CM obtained from single donors and pooled CM. We also assessed the impact of lyophilization on the immunomodulatory properties of CM.
RESULTS
We demonstrate that CM from hAM has comparable immunomodulatory properties to CM from hAMSC at passages 0 and 2. Furthermore, we demonstrate that pooled CMs have similar effects when compared to CM from single donors used separately. Finally, we demonstrate that lyophilization does not alter the in vitro immunomodulatory properties of CM from hAM and hAMSC.
CONCLUSIONS
The results presented herein support the possibility to produce secretome from intact hAM and open the prospect to highly improve the scalability of the GMP production process while reducing the costs and time related to the process of cell isolation and expansion. Moreover, the possibility of having a lyophilized secretome that maintains its original properties would allow for a ready-to-use product with easier handling, shipping and storage. The use of a lyophilized product will also facilitate clinicians by permitting customized reconstitution volumes and methods according to the most suitable formula required by the clinical application.
Identifiants
pubmed: 34641958
doi: 10.1186/s13287-021-02607-z
pii: 10.1186/s13287-021-02607-z
pmc: PMC8513276
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
540Informations de copyright
© 2021. The Author(s).
Références
Arthritis Rheumatol. 2014 Feb;66(2):327-39
pubmed: 24504805
Front Immunol. 2020 Jun 09;11:1156
pubmed: 32582218
Eplasty. 2008 Apr 11;8:e21
pubmed: 18470282
Cell Rep. 2018 Feb 27;22(9):2504-2517
pubmed: 29490284
Crit Care Med. 2016 Nov;44(11):e1118-e1131
pubmed: 27441900
Cell Transplant. 2009;18(4):405-22
pubmed: 19622228
PLoS One. 2012;7(7):e41105
pubmed: 22815931
Adv Wound Care (New Rochelle). 2016 Feb 1;5(2):65-78
pubmed: 26862464
J Cell Mol Med. 2014 Aug;18(8):1631-43
pubmed: 24894806
Placenta. 2016 Dec;48:99-103
pubmed: 27871479
Eplasty. 2008 Apr 07;8:e18
pubmed: 18461121
Int J Mol Sci. 2017 Aug 25;18(9):
pubmed: 28841158
Front Bioeng Biotechnol. 2015 Oct 19;3:162
pubmed: 26539433
Stem Cells. 2008 Jan;26(1):182-92
pubmed: 17901399
Cell Transplant. 2015;24(9):1733-52
pubmed: 25259480
Clin Ophthalmol. 2018 Jun 18;12:1105-1112
pubmed: 29950805
J Tissue Eng Regen Med. 2017 Oct;11(10):2895-2911
pubmed: 27396853
Cell Transplant. 2017 Apr 13;26(4):531-539
pubmed: 27938500
Invest Ophthalmol Vis Sci. 2005 Mar;46(3):900-7
pubmed: 15728546
PLoS One. 2012;7(10):e46956
pubmed: 23071674
Cytotherapy. 2014 Jan;16(1):17-32
pubmed: 24094500
Methods Mol Biol. 2016;1416:233-44
pubmed: 27236675
Cytotherapy. 2016 Feb;18(2):151-9
pubmed: 26724220
Cell Transplant. 2015;24(12):2601-14
pubmed: 25812083
Cytometry A. 2015 Dec;87(12):1067-9
pubmed: 26506224
Stem Cell Rev Rep. 2015 Jun;11(3):394-407
pubmed: 25348066
J Cell Mol Med. 2016 Jan;20(1):157-69
pubmed: 26515425
J Cell Mol Med. 2019 Feb;23(2):1581-1592
pubmed: 30585395
Stem Cells Transl Med. 2020 Sep;9(9):1023-1035
pubmed: 32452646
Stem Cells Transl Med. 2016 Nov;5(11):1473-1484
pubmed: 27405780
Cytotherapy. 2012 Feb;14(2):153-61
pubmed: 21954836
Curr Eye Res. 2020 Dec;45(12):1490-1496
pubmed: 32338541
Sci Rep. 2019 Oct 2;9(1):14193
pubmed: 31578445
Clin Exp Rheumatol. 2015 Jul-Aug;33(4):484-90
pubmed: 25962385
Front Cell Dev Biol. 2021 Apr 13;9:648472
pubmed: 33928083
Front Bioeng Biotechnol. 2019 Apr 10;7:75
pubmed: 31024907