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Human platelet lysate in mesenchymal stromal cell expansion according to a GMP grade protocol: a cell factory experience

BACKGROUND: The use of platelet lysate (PL) for the ex-vivo expansion of mesenchymal stromal/stem cells (MSCs) was initially proposed by Doucet et al. in 2005, as an alternative to animal serum. Moreover, regulatory authorities discourage the use of fetal bovine serum (FBS) or other animal derivativ...

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Autores principales: Becherucci, Valentina, Piccini, Luisa, Casamassima, Serena, Bisin, Silvia, Gori, Valentina, Gentile, Francesca, Ceccantini, Riccardo, De Rienzo, Elena, Bindi, Barbara, Pavan, Paola, Cunial, Vanessa, Allegro, Elisa, Ermini, Stefano, Brugnolo, Francesca, Astori, Giuseppe, Bambi, Franco
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5930506/
https://www.ncbi.nlm.nih.gov/pubmed/29720245
http://dx.doi.org/10.1186/s13287-018-0863-8
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author Becherucci, Valentina
Piccini, Luisa
Casamassima, Serena
Bisin, Silvia
Gori, Valentina
Gentile, Francesca
Ceccantini, Riccardo
De Rienzo, Elena
Bindi, Barbara
Pavan, Paola
Cunial, Vanessa
Allegro, Elisa
Ermini, Stefano
Brugnolo, Francesca
Astori, Giuseppe
Bambi, Franco
author_facet Becherucci, Valentina
Piccini, Luisa
Casamassima, Serena
Bisin, Silvia
Gori, Valentina
Gentile, Francesca
Ceccantini, Riccardo
De Rienzo, Elena
Bindi, Barbara
Pavan, Paola
Cunial, Vanessa
Allegro, Elisa
Ermini, Stefano
Brugnolo, Francesca
Astori, Giuseppe
Bambi, Franco
author_sort Becherucci, Valentina
collection PubMed
description BACKGROUND: The use of platelet lysate (PL) for the ex-vivo expansion of mesenchymal stromal/stem cells (MSCs) was initially proposed by Doucet et al. in 2005, as an alternative to animal serum. Moreover, regulatory authorities discourage the use of fetal bovine serum (FBS) or other animal derivatives, to avoid risk of zoonoses and xenogeneic immune reactions. Even if many studies investigated PL composition, there still are some open issues related to its use in ex-vivo MSC expansion, especially according to good manufacturing practice (GMP) grade protocols. METHODS: As an authorized cell factory, we report our experience using standardized PL produced by Azienda Ospedaliero Universitaria Meyer Transfusion Service for MSC expansion according to a GMP grade clinical protocol. As suggested by other authors, we performed an in-vitro test on MSCs versus MSCs cultured with FBS that still represents the best way to test PL batches. We compared 12 MSC batches cultured with DMEM 5% PL with similar batches cultured with DMEM 10% FBS, focusing on the MSC proliferation rate, MSC surface marker expression, MSC immunomodulatory and differentiation potential, and finally MSC relative telomere length. RESULTS: Results confirmed the literature data as PL increases cell proliferation without affecting the MSC immunophenotype, immunomodulatory potential, differentiation potential and relative telomere length. CONCLUSIONS: PL can be considered a safe alternative to FBS for ex-vivo expansion of MSC according to a GMP grade protocol. Our experience confirms the literature data: a large number of MSCs for clinical applications can be obtained by expansion with PL, without affecting the MSC main features. Our experience underlines the benefits of a close collaboration between the PL producers (transfusion service) and the end users (cell factory) in a synergy of skills and experiences that can lead to standardized PL production. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13287-018-0863-8) contains supplementary material, which is available to authorized users.
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spelling pubmed-59305062018-05-09 Human platelet lysate in mesenchymal stromal cell expansion according to a GMP grade protocol: a cell factory experience Becherucci, Valentina Piccini, Luisa Casamassima, Serena Bisin, Silvia Gori, Valentina Gentile, Francesca Ceccantini, Riccardo De Rienzo, Elena Bindi, Barbara Pavan, Paola Cunial, Vanessa Allegro, Elisa Ermini, Stefano Brugnolo, Francesca Astori, Giuseppe Bambi, Franco Stem Cell Res Ther Research BACKGROUND: The use of platelet lysate (PL) for the ex-vivo expansion of mesenchymal stromal/stem cells (MSCs) was initially proposed by Doucet et al. in 2005, as an alternative to animal serum. Moreover, regulatory authorities discourage the use of fetal bovine serum (FBS) or other animal derivatives, to avoid risk of zoonoses and xenogeneic immune reactions. Even if many studies investigated PL composition, there still are some open issues related to its use in ex-vivo MSC expansion, especially according to good manufacturing practice (GMP) grade protocols. METHODS: As an authorized cell factory, we report our experience using standardized PL produced by Azienda Ospedaliero Universitaria Meyer Transfusion Service for MSC expansion according to a GMP grade clinical protocol. As suggested by other authors, we performed an in-vitro test on MSCs versus MSCs cultured with FBS that still represents the best way to test PL batches. We compared 12 MSC batches cultured with DMEM 5% PL with similar batches cultured with DMEM 10% FBS, focusing on the MSC proliferation rate, MSC surface marker expression, MSC immunomodulatory and differentiation potential, and finally MSC relative telomere length. RESULTS: Results confirmed the literature data as PL increases cell proliferation without affecting the MSC immunophenotype, immunomodulatory potential, differentiation potential and relative telomere length. CONCLUSIONS: PL can be considered a safe alternative to FBS for ex-vivo expansion of MSC according to a GMP grade protocol. Our experience confirms the literature data: a large number of MSCs for clinical applications can be obtained by expansion with PL, without affecting the MSC main features. Our experience underlines the benefits of a close collaboration between the PL producers (transfusion service) and the end users (cell factory) in a synergy of skills and experiences that can lead to standardized PL production. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13287-018-0863-8) contains supplementary material, which is available to authorized users. BioMed Central 2018-05-02 /pmc/articles/PMC5930506/ /pubmed/29720245 http://dx.doi.org/10.1186/s13287-018-0863-8 Text en © The Author(s). 2018 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
Becherucci, Valentina
Piccini, Luisa
Casamassima, Serena
Bisin, Silvia
Gori, Valentina
Gentile, Francesca
Ceccantini, Riccardo
De Rienzo, Elena
Bindi, Barbara
Pavan, Paola
Cunial, Vanessa
Allegro, Elisa
Ermini, Stefano
Brugnolo, Francesca
Astori, Giuseppe
Bambi, Franco
Human platelet lysate in mesenchymal stromal cell expansion according to a GMP grade protocol: a cell factory experience
title Human platelet lysate in mesenchymal stromal cell expansion according to a GMP grade protocol: a cell factory experience
title_full Human platelet lysate in mesenchymal stromal cell expansion according to a GMP grade protocol: a cell factory experience
title_fullStr Human platelet lysate in mesenchymal stromal cell expansion according to a GMP grade protocol: a cell factory experience
title_full_unstemmed Human platelet lysate in mesenchymal stromal cell expansion according to a GMP grade protocol: a cell factory experience
title_short Human platelet lysate in mesenchymal stromal cell expansion according to a GMP grade protocol: a cell factory experience
title_sort human platelet lysate in mesenchymal stromal cell expansion according to a gmp grade protocol: a cell factory experience
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5930506/
https://www.ncbi.nlm.nih.gov/pubmed/29720245
http://dx.doi.org/10.1186/s13287-018-0863-8
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