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The production method affects the efficacy of platelet derivatives to expand mesenchymal stromal cells in vitro
BACKGROUND: The use of fetal bovine serum (FBS) as a media supplement for the ex vivo expansion of bone-marrow derived mesenchymal stromal cells (BM-MSC) has been discouraged by regulatory agencies, due to the risk of transmitting zoonoses and to elicit immune reactions in the host once transplanted...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5412035/ https://www.ncbi.nlm.nih.gov/pubmed/28460641 http://dx.doi.org/10.1186/s12967-017-1185-9 |
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author | Bernardi, Martina Agostini, Francesco Chieregato, Katia Amati, Eliana Durante, Cristina Rassu, Mario Ruggeri, Marco Sella, Sabrina Lombardi, Elisabetta Mazzucato, Mario Astori, Giuseppe |
author_facet | Bernardi, Martina Agostini, Francesco Chieregato, Katia Amati, Eliana Durante, Cristina Rassu, Mario Ruggeri, Marco Sella, Sabrina Lombardi, Elisabetta Mazzucato, Mario Astori, Giuseppe |
author_sort | Bernardi, Martina |
collection | PubMed |
description | BACKGROUND: The use of fetal bovine serum (FBS) as a media supplement for the ex vivo expansion of bone-marrow derived mesenchymal stromal cells (BM-MSC) has been discouraged by regulatory agencies, due to the risk of transmitting zoonoses and to elicit immune reactions in the host once transplanted. Platelet derivatives are valid FBS substitutes due to their content of growth factors that can be released disrupting the platelets by physical methods or physiological stimuli. We compared platelet derivatives produced by freezing/thawing (platelet lysates, PL) or after CaCl(2) activation (platelet releasate surnatant rich in growth factors, PR-SRGF) for their content in growth factors and their ability to support the ex vivo expansion of BM-MSC. METHODS: The cytokine content in the two platelet derivatives was evaluated. BM-MSC were expanded in complete medium containing 10, 7.5 and 5% PL or PR-SRGF and the cell phenotype, clonogenic capacity, immunomodulation properties and tri-lineage differentiation potential of the expanded cells in both media were investigated. RESULTS: The concentration of PDGF-AB, PDGF-AA, PDGF-BB in PR-SRGF resulted to be respectively 5.7×, 1.7× and 2.3× higher compared to PL. PR-SRGF promoted a higher BM-MSC proliferation rate compared to PL not altering BM-MSC phenotype. Colony forming efficiency of BM-MSC expanded in PR-SRGF showed a frequency of colonies significantly higher than cells expanded in PL. BM-MSC expanded in PL or PR-SRGF maintained their immunomodulatory properties against activated lymphocytes even if BM-MSC expanded in FBS performed significantly better. CONCLUSIONS: The method used to release platelet factors significantly affects the enrichment in growth factors and overall product performance. The standardization of the production process of platelet derivatives and the definition of their release criteria requires further investigation. |
format | Online Article Text |
id | pubmed-5412035 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-54120352017-05-03 The production method affects the efficacy of platelet derivatives to expand mesenchymal stromal cells in vitro Bernardi, Martina Agostini, Francesco Chieregato, Katia Amati, Eliana Durante, Cristina Rassu, Mario Ruggeri, Marco Sella, Sabrina Lombardi, Elisabetta Mazzucato, Mario Astori, Giuseppe J Transl Med Research BACKGROUND: The use of fetal bovine serum (FBS) as a media supplement for the ex vivo expansion of bone-marrow derived mesenchymal stromal cells (BM-MSC) has been discouraged by regulatory agencies, due to the risk of transmitting zoonoses and to elicit immune reactions in the host once transplanted. Platelet derivatives are valid FBS substitutes due to their content of growth factors that can be released disrupting the platelets by physical methods or physiological stimuli. We compared platelet derivatives produced by freezing/thawing (platelet lysates, PL) or after CaCl(2) activation (platelet releasate surnatant rich in growth factors, PR-SRGF) for their content in growth factors and their ability to support the ex vivo expansion of BM-MSC. METHODS: The cytokine content in the two platelet derivatives was evaluated. BM-MSC were expanded in complete medium containing 10, 7.5 and 5% PL or PR-SRGF and the cell phenotype, clonogenic capacity, immunomodulation properties and tri-lineage differentiation potential of the expanded cells in both media were investigated. RESULTS: The concentration of PDGF-AB, PDGF-AA, PDGF-BB in PR-SRGF resulted to be respectively 5.7×, 1.7× and 2.3× higher compared to PL. PR-SRGF promoted a higher BM-MSC proliferation rate compared to PL not altering BM-MSC phenotype. Colony forming efficiency of BM-MSC expanded in PR-SRGF showed a frequency of colonies significantly higher than cells expanded in PL. BM-MSC expanded in PL or PR-SRGF maintained their immunomodulatory properties against activated lymphocytes even if BM-MSC expanded in FBS performed significantly better. CONCLUSIONS: The method used to release platelet factors significantly affects the enrichment in growth factors and overall product performance. The standardization of the production process of platelet derivatives and the definition of their release criteria requires further investigation. BioMed Central 2017-05-01 /pmc/articles/PMC5412035/ /pubmed/28460641 http://dx.doi.org/10.1186/s12967-017-1185-9 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Bernardi, Martina Agostini, Francesco Chieregato, Katia Amati, Eliana Durante, Cristina Rassu, Mario Ruggeri, Marco Sella, Sabrina Lombardi, Elisabetta Mazzucato, Mario Astori, Giuseppe The production method affects the efficacy of platelet derivatives to expand mesenchymal stromal cells in vitro |
title | The production method affects the efficacy of platelet derivatives to expand mesenchymal stromal cells in vitro |
title_full | The production method affects the efficacy of platelet derivatives to expand mesenchymal stromal cells in vitro |
title_fullStr | The production method affects the efficacy of platelet derivatives to expand mesenchymal stromal cells in vitro |
title_full_unstemmed | The production method affects the efficacy of platelet derivatives to expand mesenchymal stromal cells in vitro |
title_short | The production method affects the efficacy of platelet derivatives to expand mesenchymal stromal cells in vitro |
title_sort | production method affects the efficacy of platelet derivatives to expand mesenchymal stromal cells in vitro |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5412035/ https://www.ncbi.nlm.nih.gov/pubmed/28460641 http://dx.doi.org/10.1186/s12967-017-1185-9 |
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