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Culture Condition of Bone Marrow Stromal Cells Affects Quantity and Quality of the Extracellular Vesicles

Extracellular vesicles (EVs) released by bone marrow stromal cells (BMSCs) have been shown to act as a transporter of bioactive molecules such as RNAs and proteins in the therapeutic actions of BMSCs in various diseases. Although EV therapy holds great promise to be a safer cell-free therapy overcom...

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Autores principales: Scheiber, Amanda L., Clark, Cierra A., Kaito, Takashi, Iwamoto, Masahiro, Horwitz, Edwin M., Kawasawa, Yuka Imamura, Otsuru, Satoru
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8834965/
https://www.ncbi.nlm.nih.gov/pubmed/35162938
http://dx.doi.org/10.3390/ijms23031017
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author Scheiber, Amanda L.
Clark, Cierra A.
Kaito, Takashi
Iwamoto, Masahiro
Horwitz, Edwin M.
Kawasawa, Yuka Imamura
Otsuru, Satoru
author_facet Scheiber, Amanda L.
Clark, Cierra A.
Kaito, Takashi
Iwamoto, Masahiro
Horwitz, Edwin M.
Kawasawa, Yuka Imamura
Otsuru, Satoru
author_sort Scheiber, Amanda L.
collection PubMed
description Extracellular vesicles (EVs) released by bone marrow stromal cells (BMSCs) have been shown to act as a transporter of bioactive molecules such as RNAs and proteins in the therapeutic actions of BMSCs in various diseases. Although EV therapy holds great promise to be a safer cell-free therapy overcoming issues related to cell therapy, manufacturing processes that offer scalable and reproducible EV production have not been established. Robust and scalable BMSC manufacturing methods have been shown to enhance EV production; however, the effects on EV quality remain less studied. Here, using human BMSCs isolated from nine healthy donors, we examined the effects of high-performance culture media that can rapidly expand BMSCs on EV production and quality in comparison with the conventional culture medium. We found significantly increased EV production from BMSCs cultured in the high-performance media without altering their multipotency and immunophenotypes. RNA sequencing revealed that RNA contents in EVs from high-performance media were significantly reduced with altered profiles of microRNA enriched in those related to cellular growth and proliferation in the pathway analysis. Given that pre-clinical studies at the laboratory scale often use the conventional medium, these findings could account for the discrepancy in outcomes between pre-clinical and clinical studies. Therefore, this study highlights the importance of selecting proper culture conditions for scalable and reproducible EV manufacturing.
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spelling pubmed-88349652022-02-12 Culture Condition of Bone Marrow Stromal Cells Affects Quantity and Quality of the Extracellular Vesicles Scheiber, Amanda L. Clark, Cierra A. Kaito, Takashi Iwamoto, Masahiro Horwitz, Edwin M. Kawasawa, Yuka Imamura Otsuru, Satoru Int J Mol Sci Article Extracellular vesicles (EVs) released by bone marrow stromal cells (BMSCs) have been shown to act as a transporter of bioactive molecules such as RNAs and proteins in the therapeutic actions of BMSCs in various diseases. Although EV therapy holds great promise to be a safer cell-free therapy overcoming issues related to cell therapy, manufacturing processes that offer scalable and reproducible EV production have not been established. Robust and scalable BMSC manufacturing methods have been shown to enhance EV production; however, the effects on EV quality remain less studied. Here, using human BMSCs isolated from nine healthy donors, we examined the effects of high-performance culture media that can rapidly expand BMSCs on EV production and quality in comparison with the conventional culture medium. We found significantly increased EV production from BMSCs cultured in the high-performance media without altering their multipotency and immunophenotypes. RNA sequencing revealed that RNA contents in EVs from high-performance media were significantly reduced with altered profiles of microRNA enriched in those related to cellular growth and proliferation in the pathway analysis. Given that pre-clinical studies at the laboratory scale often use the conventional medium, these findings could account for the discrepancy in outcomes between pre-clinical and clinical studies. Therefore, this study highlights the importance of selecting proper culture conditions for scalable and reproducible EV manufacturing. MDPI 2022-01-18 /pmc/articles/PMC8834965/ /pubmed/35162938 http://dx.doi.org/10.3390/ijms23031017 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
Scheiber, Amanda L.
Clark, Cierra A.
Kaito, Takashi
Iwamoto, Masahiro
Horwitz, Edwin M.
Kawasawa, Yuka Imamura
Otsuru, Satoru
Culture Condition of Bone Marrow Stromal Cells Affects Quantity and Quality of the Extracellular Vesicles
title Culture Condition of Bone Marrow Stromal Cells Affects Quantity and Quality of the Extracellular Vesicles
title_full Culture Condition of Bone Marrow Stromal Cells Affects Quantity and Quality of the Extracellular Vesicles
title_fullStr Culture Condition of Bone Marrow Stromal Cells Affects Quantity and Quality of the Extracellular Vesicles
title_full_unstemmed Culture Condition of Bone Marrow Stromal Cells Affects Quantity and Quality of the Extracellular Vesicles
title_short Culture Condition of Bone Marrow Stromal Cells Affects Quantity and Quality of the Extracellular Vesicles
title_sort culture condition of bone marrow stromal cells affects quantity and quality of the extracellular vesicles
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8834965/
https://www.ncbi.nlm.nih.gov/pubmed/35162938
http://dx.doi.org/10.3390/ijms23031017
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