Exosomes From Human Cardiac Progenitor Cells for Therapeutic Applications: Development of a GMP-Grade Manufacturing Method
Exosomes, nanosized membrane vesicles secreted by cardiac progenitor cells (Exo-CPC), inhibit cardiomyocyte apoptosis under stress conditions, promote angiogenesis in vitro, and prevent the early decline in cardiac function after myocardial infarction in vivo in preclinical rat models. The recogniti...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2018-08-01
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| Series: | Frontiers in Physiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/article/10.3389/fphys.2018.01169/full |
| _version_ | 1819071229855793152 |
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| author | Gabriella Andriolo Gabriella Andriolo Elena Provasi Elena Provasi Viviana Lo Cicero Viviana Lo Cicero Andrea Brambilla Andrea Brambilla Sabrina Soncin Sabrina Soncin Tiziano Torre Giuseppina Milano Giuseppina Milano Giuseppina Milano Vanessa Biemmi Vanessa Biemmi Giuseppe Vassalli Giuseppe Vassalli Giuseppe Vassalli Lucia Turchetto Lucia Turchetto Lucio Barile Lucio Barile Marina Radrizzani Marina Radrizzani |
| author_facet | Gabriella Andriolo Gabriella Andriolo Elena Provasi Elena Provasi Viviana Lo Cicero Viviana Lo Cicero Andrea Brambilla Andrea Brambilla Sabrina Soncin Sabrina Soncin Tiziano Torre Giuseppina Milano Giuseppina Milano Giuseppina Milano Vanessa Biemmi Vanessa Biemmi Giuseppe Vassalli Giuseppe Vassalli Giuseppe Vassalli Lucia Turchetto Lucia Turchetto Lucio Barile Lucio Barile Marina Radrizzani Marina Radrizzani |
| author_sort | Gabriella Andriolo |
| collection | DOAJ |
| description | Exosomes, nanosized membrane vesicles secreted by cardiac progenitor cells (Exo-CPC), inhibit cardiomyocyte apoptosis under stress conditions, promote angiogenesis in vitro, and prevent the early decline in cardiac function after myocardial infarction in vivo in preclinical rat models. The recognition of exosome-mediated effects has moved attempts at developing cell-free approaches for cardiac repair. Such approaches offer major advantages including the fact that exosomes can be stored as ready-to-use agents and delivered to patients with acute coronary syndromes. The aim of the present work was the development of a good manufacturing practice (GMP)-grade method for the large-scale preparation of Exo-CPC as a medicinal product, for a future clinical translation. A GMP-compliant manufacturing method was set up, based on large-scale cell culture in xeno-free conditions, collection of up to 8 l of exosome-containing conditioned medium and isolation of Exo-CPC through tangential flow filtration. Quality control tests were developed and carried out to evaluate safety, identity, and potency of both cardiac progenitor cells (CPC) as cell source and Exo-CPC as final product (GMP-Exo-CPC). CPC, cultured in xeno-free conditions, showed a lower doubling-time than observed in research-grade condition, while producing exosomes with similar features. Cells showed the typical phenotype of mesenchymal progenitor cells (CD73/CD90/CD105 positive, CD14/CD20/CD34/CD45/HLA-DR negative), and expressed mesodermal (TBX5/TBX18) and cardiac-specific (GATA4/MESP1) transcription factors. Purified GMP-Exo-CPC showed the typical nanoparticle tracking analysis profile and expressed main exosome markers (CD9/CD63/CD81/TSG101). The GMP manufacturing method guaranteed high exosome yield (>1013 particles) and consistent removal (≥97%) of contaminating proteins. The resulting GMP-Exo-CPC were tested for safety, purity, identity, and potency in vitro, showing functional anti-apoptotic and pro-angiogenic activity. The therapeutic efficacy was validated in vivo in rats, where GMP-Exo-CPC ameliorated heart function after myocardial infarction. Our standardized production method and testing strategy for large-scale manufacturing of GMP-Exo-CPC open new perspectives for reliable human therapeutic applications for acute myocardial infarction syndrome and can be easily applied to other cell sources for different therapeutic areas. |
| first_indexed | 2024-12-21T17:18:31Z |
| format | Article |
| id | doaj.art-c227e996d7f544468a8d380a3c1c0e39 |
| institution | Directory Open Access Journal |
| issn | 1664-042X |
| language | English |
| last_indexed | 2024-12-21T17:18:31Z |
| publishDate | 2018-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Physiology |
| spelling | doaj.art-c227e996d7f544468a8d380a3c1c0e392022-12-21T18:56:13ZengFrontiers Media S.A.Frontiers in Physiology1664-042X2018-08-01910.3389/fphys.2018.01169400470Exosomes From Human Cardiac Progenitor Cells for Therapeutic Applications: Development of a GMP-Grade Manufacturing MethodGabriella Andriolo0Gabriella Andriolo1Elena Provasi2Elena Provasi3Viviana Lo Cicero4Viviana Lo Cicero5Andrea Brambilla6Andrea Brambilla7Sabrina Soncin8Sabrina Soncin9Tiziano Torre10Giuseppina Milano11Giuseppina Milano12Giuseppina Milano13Vanessa Biemmi14Vanessa Biemmi15Giuseppe Vassalli16Giuseppe Vassalli17Giuseppe Vassalli18Lucia Turchetto19Lucia Turchetto20Lucio Barile21Lucio Barile22Marina Radrizzani23Marina Radrizzani24Lugano Cell Factory, Fondazione Cardiocentro Ticino, Lugano, SwitzerlandSwiss Institute for Regenerative Medicine, Torricella-Taverne, SwitzerlandLugano Cell Factory, Fondazione Cardiocentro Ticino, Lugano, SwitzerlandSwiss Institute for Regenerative Medicine, Torricella-Taverne, SwitzerlandLugano Cell Factory, Fondazione Cardiocentro Ticino, Lugano, SwitzerlandSwiss Institute for Regenerative Medicine, Torricella-Taverne, SwitzerlandLugano Cell Factory, Fondazione Cardiocentro Ticino, Lugano, SwitzerlandSwiss Institute for Regenerative Medicine, Torricella-Taverne, SwitzerlandLugano Cell Factory, Fondazione Cardiocentro Ticino, Lugano, SwitzerlandSwiss Institute for Regenerative Medicine, Torricella-Taverne, SwitzerlandDivision of Cardiac Surgery, Fondazione Cardiocentro Ticino, Lugano, SwitzerlandSwiss Institute for Regenerative Medicine, Torricella-Taverne, SwitzerlandLaboratory of Molecular and Cellular Cardiology, Fondazione Cardiocentro Ticino, Lugano, SwitzerlandDepartment of Heart and Vessels, Laboratory of Cardiovascular Research, Lausanne University Hospital, Lausanne, SwitzerlandSwiss Institute for Regenerative Medicine, Torricella-Taverne, SwitzerlandLaboratory of Molecular and Cellular Cardiology, Fondazione Cardiocentro Ticino, Lugano, SwitzerlandSwiss Institute for Regenerative Medicine, Torricella-Taverne, SwitzerlandLaboratory of Molecular and Cellular Cardiology, Fondazione Cardiocentro Ticino, Lugano, SwitzerlandDivision of Cardiology, Fondazione Cardiocentro Ticino, Lugano, SwitzerlandLugano Cell Factory, Fondazione Cardiocentro Ticino, Lugano, SwitzerlandSwiss Institute for Regenerative Medicine, Torricella-Taverne, SwitzerlandSwiss Institute for Regenerative Medicine, Torricella-Taverne, SwitzerlandLaboratory of Molecular and Cellular Cardiology, Fondazione Cardiocentro Ticino, Lugano, SwitzerlandLugano Cell Factory, Fondazione Cardiocentro Ticino, Lugano, SwitzerlandSwiss Institute for Regenerative Medicine, Torricella-Taverne, SwitzerlandExosomes, nanosized membrane vesicles secreted by cardiac progenitor cells (Exo-CPC), inhibit cardiomyocyte apoptosis under stress conditions, promote angiogenesis in vitro, and prevent the early decline in cardiac function after myocardial infarction in vivo in preclinical rat models. The recognition of exosome-mediated effects has moved attempts at developing cell-free approaches for cardiac repair. Such approaches offer major advantages including the fact that exosomes can be stored as ready-to-use agents and delivered to patients with acute coronary syndromes. The aim of the present work was the development of a good manufacturing practice (GMP)-grade method for the large-scale preparation of Exo-CPC as a medicinal product, for a future clinical translation. A GMP-compliant manufacturing method was set up, based on large-scale cell culture in xeno-free conditions, collection of up to 8 l of exosome-containing conditioned medium and isolation of Exo-CPC through tangential flow filtration. Quality control tests were developed and carried out to evaluate safety, identity, and potency of both cardiac progenitor cells (CPC) as cell source and Exo-CPC as final product (GMP-Exo-CPC). CPC, cultured in xeno-free conditions, showed a lower doubling-time than observed in research-grade condition, while producing exosomes with similar features. Cells showed the typical phenotype of mesenchymal progenitor cells (CD73/CD90/CD105 positive, CD14/CD20/CD34/CD45/HLA-DR negative), and expressed mesodermal (TBX5/TBX18) and cardiac-specific (GATA4/MESP1) transcription factors. Purified GMP-Exo-CPC showed the typical nanoparticle tracking analysis profile and expressed main exosome markers (CD9/CD63/CD81/TSG101). The GMP manufacturing method guaranteed high exosome yield (>1013 particles) and consistent removal (≥97%) of contaminating proteins. The resulting GMP-Exo-CPC were tested for safety, purity, identity, and potency in vitro, showing functional anti-apoptotic and pro-angiogenic activity. The therapeutic efficacy was validated in vivo in rats, where GMP-Exo-CPC ameliorated heart function after myocardial infarction. Our standardized production method and testing strategy for large-scale manufacturing of GMP-Exo-CPC open new perspectives for reliable human therapeutic applications for acute myocardial infarction syndrome and can be easily applied to other cell sources for different therapeutic areas.https://www.frontiersin.org/article/10.3389/fphys.2018.01169/fullexosomescardiac progenitor cellsgood manufacturing practiceslarge-scale productionquality control |
| spellingShingle | Gabriella Andriolo Gabriella Andriolo Elena Provasi Elena Provasi Viviana Lo Cicero Viviana Lo Cicero Andrea Brambilla Andrea Brambilla Sabrina Soncin Sabrina Soncin Tiziano Torre Giuseppina Milano Giuseppina Milano Giuseppina Milano Vanessa Biemmi Vanessa Biemmi Giuseppe Vassalli Giuseppe Vassalli Giuseppe Vassalli Lucia Turchetto Lucia Turchetto Lucio Barile Lucio Barile Marina Radrizzani Marina Radrizzani Exosomes From Human Cardiac Progenitor Cells for Therapeutic Applications: Development of a GMP-Grade Manufacturing Method Frontiers in Physiology exosomes cardiac progenitor cells good manufacturing practices large-scale production quality control |
| title | Exosomes From Human Cardiac Progenitor Cells for Therapeutic Applications: Development of a GMP-Grade Manufacturing Method |
| title_full | Exosomes From Human Cardiac Progenitor Cells for Therapeutic Applications: Development of a GMP-Grade Manufacturing Method |
| title_fullStr | Exosomes From Human Cardiac Progenitor Cells for Therapeutic Applications: Development of a GMP-Grade Manufacturing Method |
| title_full_unstemmed | Exosomes From Human Cardiac Progenitor Cells for Therapeutic Applications: Development of a GMP-Grade Manufacturing Method |
| title_short | Exosomes From Human Cardiac Progenitor Cells for Therapeutic Applications: Development of a GMP-Grade Manufacturing Method |
| title_sort | exosomes from human cardiac progenitor cells for therapeutic applications development of a gmp grade manufacturing method |
| topic | exosomes cardiac progenitor cells good manufacturing practices large-scale production quality control |
| url | https://www.frontiersin.org/article/10.3389/fphys.2018.01169/full |
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