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Freeze-Dried Mesenchymal Stem Cell-Secretome Pharmaceuticalization: Optimization of Formulation and Manufacturing Process Robustness

Producing mesenchymal stem cell (MSC)-secretome for dose escalation studies and clinical practice requires scalable and good manufacturing practice (GMP)-compliant production procedures and formulation into a standardized medicinal product. Starting from a method that combines ultrafiltration and fr...

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Autores principales: Mocchi, Michela, Bari, Elia, Marrubini, Giorgio, Bonda, Andrea Foglio, Perteghella, Sara, Tartara, Fulvio, Cofano, Fabio, Perna, Giuseppe di, Giovannelli, Lorella, Mandracchia, Delia, Sorlini, Marzio, Garbossa, Diego, Torre, Maria Luisa, Segale, Lorena
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8401234/
https://www.ncbi.nlm.nih.gov/pubmed/34452088
http://dx.doi.org/10.3390/pharmaceutics13081129
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author Mocchi, Michela
Bari, Elia
Marrubini, Giorgio
Bonda, Andrea Foglio
Perteghella, Sara
Tartara, Fulvio
Cofano, Fabio
Perna, Giuseppe di
Giovannelli, Lorella
Mandracchia, Delia
Sorlini, Marzio
Garbossa, Diego
Torre, Maria Luisa
Segale, Lorena
author_facet Mocchi, Michela
Bari, Elia
Marrubini, Giorgio
Bonda, Andrea Foglio
Perteghella, Sara
Tartara, Fulvio
Cofano, Fabio
Perna, Giuseppe di
Giovannelli, Lorella
Mandracchia, Delia
Sorlini, Marzio
Garbossa, Diego
Torre, Maria Luisa
Segale, Lorena
author_sort Mocchi, Michela
collection PubMed
description Producing mesenchymal stem cell (MSC)-secretome for dose escalation studies and clinical practice requires scalable and good manufacturing practice (GMP)-compliant production procedures and formulation into a standardized medicinal product. Starting from a method that combines ultrafiltration and freeze-drying to transform MSC-secretome into a pharmaceutical product, the lyosecretome, this work aims to: (i) optimize the lyosecretome formulation; (ii) investigate sources of variability that can affect the robustness of the manufacturing process; (iii) modify the ultrafiltration step to obtain a more standardized final product. Design of experiments and principal component analysis of the data were used to study the influence of batch production, lyophilization, mannitol (M)/sucrose (S) binary mixture, selected as cryoprotectant excipients, and the total amount of excipients on the extracellular vesicles (EV) particle size, the protein and lipid content and the in vitro anti-elastase. The different excipients ratios did not affect residual moisture or EV particle size; simultaneously, proteins and lipids were better preserved in the freeze-dried product using the maximum total concentration of excipients (1.5% w/v) with a M:S ratio of about 60% w/w. The anti-elastase activity was instead better preserved using 0.5% w/w of M as excipient. The secretome batch showed to be the primary source of variability; therefore, the manufacturing process has been modified and then validated: the final product is now concentrated to reach a specific protein (and lipid) concentration instead of cell equivalent concentration. The new standardization approach led to a final product with more reproducible quali-quantitative composition and higher biological activity.
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spelling pubmed-84012342021-08-29 Freeze-Dried Mesenchymal Stem Cell-Secretome Pharmaceuticalization: Optimization of Formulation and Manufacturing Process Robustness Mocchi, Michela Bari, Elia Marrubini, Giorgio Bonda, Andrea Foglio Perteghella, Sara Tartara, Fulvio Cofano, Fabio Perna, Giuseppe di Giovannelli, Lorella Mandracchia, Delia Sorlini, Marzio Garbossa, Diego Torre, Maria Luisa Segale, Lorena Pharmaceutics Article Producing mesenchymal stem cell (MSC)-secretome for dose escalation studies and clinical practice requires scalable and good manufacturing practice (GMP)-compliant production procedures and formulation into a standardized medicinal product. Starting from a method that combines ultrafiltration and freeze-drying to transform MSC-secretome into a pharmaceutical product, the lyosecretome, this work aims to: (i) optimize the lyosecretome formulation; (ii) investigate sources of variability that can affect the robustness of the manufacturing process; (iii) modify the ultrafiltration step to obtain a more standardized final product. Design of experiments and principal component analysis of the data were used to study the influence of batch production, lyophilization, mannitol (M)/sucrose (S) binary mixture, selected as cryoprotectant excipients, and the total amount of excipients on the extracellular vesicles (EV) particle size, the protein and lipid content and the in vitro anti-elastase. The different excipients ratios did not affect residual moisture or EV particle size; simultaneously, proteins and lipids were better preserved in the freeze-dried product using the maximum total concentration of excipients (1.5% w/v) with a M:S ratio of about 60% w/w. The anti-elastase activity was instead better preserved using 0.5% w/w of M as excipient. The secretome batch showed to be the primary source of variability; therefore, the manufacturing process has been modified and then validated: the final product is now concentrated to reach a specific protein (and lipid) concentration instead of cell equivalent concentration. The new standardization approach led to a final product with more reproducible quali-quantitative composition and higher biological activity. MDPI 2021-07-23 /pmc/articles/PMC8401234/ /pubmed/34452088 http://dx.doi.org/10.3390/pharmaceutics13081129 Text en © 2021 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
Mocchi, Michela
Bari, Elia
Marrubini, Giorgio
Bonda, Andrea Foglio
Perteghella, Sara
Tartara, Fulvio
Cofano, Fabio
Perna, Giuseppe di
Giovannelli, Lorella
Mandracchia, Delia
Sorlini, Marzio
Garbossa, Diego
Torre, Maria Luisa
Segale, Lorena
Freeze-Dried Mesenchymal Stem Cell-Secretome Pharmaceuticalization: Optimization of Formulation and Manufacturing Process Robustness
title Freeze-Dried Mesenchymal Stem Cell-Secretome Pharmaceuticalization: Optimization of Formulation and Manufacturing Process Robustness
title_full Freeze-Dried Mesenchymal Stem Cell-Secretome Pharmaceuticalization: Optimization of Formulation and Manufacturing Process Robustness
title_fullStr Freeze-Dried Mesenchymal Stem Cell-Secretome Pharmaceuticalization: Optimization of Formulation and Manufacturing Process Robustness
title_full_unstemmed Freeze-Dried Mesenchymal Stem Cell-Secretome Pharmaceuticalization: Optimization of Formulation and Manufacturing Process Robustness
title_short Freeze-Dried Mesenchymal Stem Cell-Secretome Pharmaceuticalization: Optimization of Formulation and Manufacturing Process Robustness
title_sort freeze-dried mesenchymal stem cell-secretome pharmaceuticalization: optimization of formulation and manufacturing process robustness
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8401234/
https://www.ncbi.nlm.nih.gov/pubmed/34452088
http://dx.doi.org/10.3390/pharmaceutics13081129
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