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A New Human Platelet Lysate for Mesenchymal Stem Cell Production Compliant with Good Manufacturing Practice Conditions

Mesenchymal stem cells (MSCs) are classified as advanced therapy medicinal products, a new category of GMP (good manufacturing practice)-compliant medicines for clinical use. We isolated MSCs from 5 bone marrow (BM) samples using human platelet lysate (HPL) instead of foetal bovine serum (FBS). We u...

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Autores principales: Mareschi, Katia, Marini, Elena, Niclot, Alessia Giovanna Santa Banche, Barone, Marta, Pinnetta, Giuseppe, Adamini, Aloe, Spadea, Manuela, Labanca, Luciana, Lucania, Graziella, Ferrero, Ivana, Fagioli, Franca
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8953582/
https://www.ncbi.nlm.nih.gov/pubmed/35328655
http://dx.doi.org/10.3390/ijms23063234
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author Mareschi, Katia
Marini, Elena
Niclot, Alessia Giovanna Santa Banche
Barone, Marta
Pinnetta, Giuseppe
Adamini, Aloe
Spadea, Manuela
Labanca, Luciana
Lucania, Graziella
Ferrero, Ivana
Fagioli, Franca
author_facet Mareschi, Katia
Marini, Elena
Niclot, Alessia Giovanna Santa Banche
Barone, Marta
Pinnetta, Giuseppe
Adamini, Aloe
Spadea, Manuela
Labanca, Luciana
Lucania, Graziella
Ferrero, Ivana
Fagioli, Franca
author_sort Mareschi, Katia
collection PubMed
description Mesenchymal stem cells (MSCs) are classified as advanced therapy medicinal products, a new category of GMP (good manufacturing practice)-compliant medicines for clinical use. We isolated MSCs from 5 bone marrow (BM) samples using human platelet lysate (HPL) instead of foetal bovine serum (FBS). We used a new method of HPL production consisting of treating platelet (PLTs) pools with Ca-Gluconate to form a gel clot, then mechanically squeezing to release growth factors. We compared the new HPL (HPL-S) with the standard (HPL-E) obtained by freezing/thawing cycles and by adding heparin. HPL-S had not PLTs and fibrinogen but the quantity of proteins and growth factors was comparable to HPL-E. Therefore, HPL-S needed fewer production steps to be in compliance with GMP conditions. The number of colonies forming unit-fibroblasts (CFU-F) and the maintenance of stem markers showed no significant differences between MSCs with HPL-E and HPL-S. The cumulative population doubling was higher in MSCs with HPL-E in the earlier passages, but we observed an inverted trend of cell growth at the fourth passage. Immunophenotypic analysis showed a significant lower expression of HLA-DR in the MSCs with HPL-S (1.30%) than HPL-E (14.10%). In conclusion, we demonstrated that HPL-S is an effective alternative for MSC production under GMP conditions.
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spelling pubmed-89535822022-03-26 A New Human Platelet Lysate for Mesenchymal Stem Cell Production Compliant with Good Manufacturing Practice Conditions Mareschi, Katia Marini, Elena Niclot, Alessia Giovanna Santa Banche Barone, Marta Pinnetta, Giuseppe Adamini, Aloe Spadea, Manuela Labanca, Luciana Lucania, Graziella Ferrero, Ivana Fagioli, Franca Int J Mol Sci Article Mesenchymal stem cells (MSCs) are classified as advanced therapy medicinal products, a new category of GMP (good manufacturing practice)-compliant medicines for clinical use. We isolated MSCs from 5 bone marrow (BM) samples using human platelet lysate (HPL) instead of foetal bovine serum (FBS). We used a new method of HPL production consisting of treating platelet (PLTs) pools with Ca-Gluconate to form a gel clot, then mechanically squeezing to release growth factors. We compared the new HPL (HPL-S) with the standard (HPL-E) obtained by freezing/thawing cycles and by adding heparin. HPL-S had not PLTs and fibrinogen but the quantity of proteins and growth factors was comparable to HPL-E. Therefore, HPL-S needed fewer production steps to be in compliance with GMP conditions. The number of colonies forming unit-fibroblasts (CFU-F) and the maintenance of stem markers showed no significant differences between MSCs with HPL-E and HPL-S. The cumulative population doubling was higher in MSCs with HPL-E in the earlier passages, but we observed an inverted trend of cell growth at the fourth passage. Immunophenotypic analysis showed a significant lower expression of HLA-DR in the MSCs with HPL-S (1.30%) than HPL-E (14.10%). In conclusion, we demonstrated that HPL-S is an effective alternative for MSC production under GMP conditions. MDPI 2022-03-17 /pmc/articles/PMC8953582/ /pubmed/35328655 http://dx.doi.org/10.3390/ijms23063234 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Mareschi, Katia
Marini, Elena
Niclot, Alessia Giovanna Santa Banche
Barone, Marta
Pinnetta, Giuseppe
Adamini, Aloe
Spadea, Manuela
Labanca, Luciana
Lucania, Graziella
Ferrero, Ivana
Fagioli, Franca
A New Human Platelet Lysate for Mesenchymal Stem Cell Production Compliant with Good Manufacturing Practice Conditions
title A New Human Platelet Lysate for Mesenchymal Stem Cell Production Compliant with Good Manufacturing Practice Conditions
title_full A New Human Platelet Lysate for Mesenchymal Stem Cell Production Compliant with Good Manufacturing Practice Conditions
title_fullStr A New Human Platelet Lysate for Mesenchymal Stem Cell Production Compliant with Good Manufacturing Practice Conditions
title_full_unstemmed A New Human Platelet Lysate for Mesenchymal Stem Cell Production Compliant with Good Manufacturing Practice Conditions
title_short A New Human Platelet Lysate for Mesenchymal Stem Cell Production Compliant with Good Manufacturing Practice Conditions
title_sort new human platelet lysate for mesenchymal stem cell production compliant with good manufacturing practice conditions
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8953582/
https://www.ncbi.nlm.nih.gov/pubmed/35328655
http://dx.doi.org/10.3390/ijms23063234
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