Exosomes From Human Cardiac Progenitor Cells for Therapeutic Applications: Development of a GMP-Grade Manufacturing Method.

Details

Serval ID
serval:BIB_1F8EB6658825
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Exosomes From Human Cardiac Progenitor Cells for Therapeutic Applications: Development of a GMP-Grade Manufacturing Method.
Journal
Frontiers in physiology
Author(s)
Andriolo G., Provasi E., Lo Cicero V., Brambilla A., Soncin S., Torre T., Milano G., Biemmi V., Vassalli G., Turchetto L., Barile L., Radrizzani M.
ISSN
1664-042X (Print)
ISSN-L
1664-042X
Publication state
Published
Issued date
2018
Peer-reviewed
Oui
Volume
9
Pages
1169
Language
english
Notes
Publication types: Journal Article
Publication Status: epublish
Abstract
Exosomes, nanosized membrane vesicles secreted by cardiac progenitor cells (Exo-CPC), inhibit cardiomyocyte apoptosis under stress conditions, promote angiogenesis <i>in vitro</i> , and prevent the early decline in cardiac function after myocardial infarction <i>in vivo</i> 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 (>10 <sup>13</sup> particles) and consistent removal (≥97%) of contaminating proteins. The resulting GMP-Exo-CPC were tested for safety, purity, identity, and potency <i>in vitro</i> , showing functional anti-apoptotic and pro-angiogenic activity. The therapeutic efficacy was validated <i>in vivo</i> 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.
Keywords
cardiac progenitor cells, exosomes, good manufacturing practices, large-scale production, quality control
Pubmed
Web of science
Open Access
Yes
Create date
17/09/2018 15:12
Last modification date
20/08/2019 13:55
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