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A Three-Dimensional Xeno-Free Culture Condition for Wharton’s Jelly-Mesenchymal Stem Cells: The Pros and Cons

Xeno-free three-dimensional cultures are gaining attention for mesenchymal stem cell (MSCs) expansion in clinical applications. We investigated the potential of xeno-free serum alternatives, human serum and human platelet lysate, to replace the current conventional use of foetal bovine serum for sub...

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Autores principales: Koh, Benson, Sulaiman, Nadiah, Fauzi, Mh Busra, Law, Jia Xian, Ng, Min Hwei, Yuan, Too Lih, Azurah, Abdul Ghani Nur, Mohd Yunus, Mohd Heikal, Idrus, Ruszymah Bt Hj, Yazid, Muhammad Dain
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9960744/
https://www.ncbi.nlm.nih.gov/pubmed/36835154
http://dx.doi.org/10.3390/ijms24043745
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author Koh, Benson
Sulaiman, Nadiah
Fauzi, Mh Busra
Law, Jia Xian
Ng, Min Hwei
Yuan, Too Lih
Azurah, Abdul Ghani Nur
Mohd Yunus, Mohd Heikal
Idrus, Ruszymah Bt Hj
Yazid, Muhammad Dain
author_facet Koh, Benson
Sulaiman, Nadiah
Fauzi, Mh Busra
Law, Jia Xian
Ng, Min Hwei
Yuan, Too Lih
Azurah, Abdul Ghani Nur
Mohd Yunus, Mohd Heikal
Idrus, Ruszymah Bt Hj
Yazid, Muhammad Dain
author_sort Koh, Benson
collection PubMed
description Xeno-free three-dimensional cultures are gaining attention for mesenchymal stem cell (MSCs) expansion in clinical applications. We investigated the potential of xeno-free serum alternatives, human serum and human platelet lysate, to replace the current conventional use of foetal bovine serum for subsequent MSCs microcarrier cultures. In this study, Wharton’s Jelly MSCs were cultured in nine different media combinations to identify the best xeno-free culture media for MSCs culture. Cell proliferation and viability were identified, and the cultured MSCs were characterised in accordance with the minimal criteria for defining multipotent mesenchymal stromal cells by the International Society for Cellular Therapy (ISCT). The selected culture media was then used in the microcarrier culture of MSCs to determine the potential of a three-dimensional culture system in the expansion of MSCs for future clinical applications, and to identify the immunomodulatory potential of cultured MSCs. Low Glucose DMEM (LG) + Human Platelet (HPL) lysate media appeared to be good candidates for replacing conventional MSCs culture media in our monolayer culture system. MSCs cultured in LG-HPL achieved high cell yield, with characteristics that remained as described by ISCT, although the overall mitochondrial activity of the cells was lower than the control and the subsequent effects remained unknown. MSC microcarrier culture, on the other hand, showed comparable cell characteristics with monolayer culture, yet had stagnated cell proliferation, which is potentially due to the inactivation of FAK. Nonetheless, both the MSCs monolayer culture and the microcarrier culture showed high suppressive activity on TNF-α, and only the MSC microcarrier culture has a better suppression of IL-1 secretion. In conclusion, LG-HPL was identified as a good xeno-free media for WJMSCs culture, and although further mechanistic research is needed, the results show that the xeno-free three-dimensional culture maintained MSC characteristics and improved immunomodulatory activities, suggesting the potential of translating the monolayer culture into this culture system in MSC expansion for future clinical application.
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spelling pubmed-99607442023-02-26 A Three-Dimensional Xeno-Free Culture Condition for Wharton’s Jelly-Mesenchymal Stem Cells: The Pros and Cons Koh, Benson Sulaiman, Nadiah Fauzi, Mh Busra Law, Jia Xian Ng, Min Hwei Yuan, Too Lih Azurah, Abdul Ghani Nur Mohd Yunus, Mohd Heikal Idrus, Ruszymah Bt Hj Yazid, Muhammad Dain Int J Mol Sci Article Xeno-free three-dimensional cultures are gaining attention for mesenchymal stem cell (MSCs) expansion in clinical applications. We investigated the potential of xeno-free serum alternatives, human serum and human platelet lysate, to replace the current conventional use of foetal bovine serum for subsequent MSCs microcarrier cultures. In this study, Wharton’s Jelly MSCs were cultured in nine different media combinations to identify the best xeno-free culture media for MSCs culture. Cell proliferation and viability were identified, and the cultured MSCs were characterised in accordance with the minimal criteria for defining multipotent mesenchymal stromal cells by the International Society for Cellular Therapy (ISCT). The selected culture media was then used in the microcarrier culture of MSCs to determine the potential of a three-dimensional culture system in the expansion of MSCs for future clinical applications, and to identify the immunomodulatory potential of cultured MSCs. Low Glucose DMEM (LG) + Human Platelet (HPL) lysate media appeared to be good candidates for replacing conventional MSCs culture media in our monolayer culture system. MSCs cultured in LG-HPL achieved high cell yield, with characteristics that remained as described by ISCT, although the overall mitochondrial activity of the cells was lower than the control and the subsequent effects remained unknown. MSC microcarrier culture, on the other hand, showed comparable cell characteristics with monolayer culture, yet had stagnated cell proliferation, which is potentially due to the inactivation of FAK. Nonetheless, both the MSCs monolayer culture and the microcarrier culture showed high suppressive activity on TNF-α, and only the MSC microcarrier culture has a better suppression of IL-1 secretion. In conclusion, LG-HPL was identified as a good xeno-free media for WJMSCs culture, and although further mechanistic research is needed, the results show that the xeno-free three-dimensional culture maintained MSC characteristics and improved immunomodulatory activities, suggesting the potential of translating the monolayer culture into this culture system in MSC expansion for future clinical application. MDPI 2023-02-13 /pmc/articles/PMC9960744/ /pubmed/36835154 http://dx.doi.org/10.3390/ijms24043745 Text en © 2023 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
Koh, Benson
Sulaiman, Nadiah
Fauzi, Mh Busra
Law, Jia Xian
Ng, Min Hwei
Yuan, Too Lih
Azurah, Abdul Ghani Nur
Mohd Yunus, Mohd Heikal
Idrus, Ruszymah Bt Hj
Yazid, Muhammad Dain
A Three-Dimensional Xeno-Free Culture Condition for Wharton’s Jelly-Mesenchymal Stem Cells: The Pros and Cons
title A Three-Dimensional Xeno-Free Culture Condition for Wharton’s Jelly-Mesenchymal Stem Cells: The Pros and Cons
title_full A Three-Dimensional Xeno-Free Culture Condition for Wharton’s Jelly-Mesenchymal Stem Cells: The Pros and Cons
title_fullStr A Three-Dimensional Xeno-Free Culture Condition for Wharton’s Jelly-Mesenchymal Stem Cells: The Pros and Cons
title_full_unstemmed A Three-Dimensional Xeno-Free Culture Condition for Wharton’s Jelly-Mesenchymal Stem Cells: The Pros and Cons
title_short A Three-Dimensional Xeno-Free Culture Condition for Wharton’s Jelly-Mesenchymal Stem Cells: The Pros and Cons
title_sort three-dimensional xeno-free culture condition for wharton’s jelly-mesenchymal stem cells: the pros and cons
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9960744/
https://www.ncbi.nlm.nih.gov/pubmed/36835154
http://dx.doi.org/10.3390/ijms24043745
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