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Effect of the Addition Frequency of 5-Azacytidine in Both Micro- and Macroscale Cultures
INTRODUCTION: Human mesenchymal stem cells (hMSCs) have a great clinical potential for tissue regeneration purposes due to its multilineage capability. Previous studies have reported that a single addition of 5-azacytidine (5-AzaC) causes the differentiation of hMSCs towards a myocardial lineage. Th...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer International Publishing
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7878657/ https://www.ncbi.nlm.nih.gov/pubmed/33633814 http://dx.doi.org/10.1007/s12195-020-00654-9 |
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author | Kadekar, Sandeep Barbe, Laurent Stoddart, Martin Varghese, Oommen P. Tenje, Maria Mestres, Gemma |
author_facet | Kadekar, Sandeep Barbe, Laurent Stoddart, Martin Varghese, Oommen P. Tenje, Maria Mestres, Gemma |
author_sort | Kadekar, Sandeep |
collection | PubMed |
description | INTRODUCTION: Human mesenchymal stem cells (hMSCs) have a great clinical potential for tissue regeneration purposes due to its multilineage capability. Previous studies have reported that a single addition of 5-azacytidine (5-AzaC) causes the differentiation of hMSCs towards a myocardial lineage. The aim of this work was to evaluate the effect of 5-AzaC addition frequency on hMSCs priming (i.e., indicating an early genetic differentiation) using two culture environments. METHODS: hMSCs were supplemented with 5-AzaC while cultured in well plates and in microfluidic chips. The impact of 5-AzaC concentration (10 and 20 μM) and addition frequency (once, daily or continuously), as well as of culture period (2 or 5 days) on the genetic upregulation of PPARγ (adipocytes), PAX3 (myoblasts), SOX9 (chondrocytes) and RUNX2 (osteoblasts) was evaluated. RESULTS: Daily delivering 5-AzaC caused a higher upregulation of PPARγ, SOX9 and RUNX2 in comparison to a single dose delivery, both under static well plates and dynamic microfluidic cultures. A particularly high gene expression of PPARγ (tenfold-change) could indicate priming of hMSCs towards adipocytes. CONCLUSIONS: Both macro- and microscale cultures provided results with similar trends, where addition frequency of 5-AzaC was a crucial factor to upregulate several genes. Microfluidics technology was proven to be a suitable platform for the continuous delivery of a drug and could be used for screening purposes in tissue engineering research. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s12195-020-00654-9) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-7878657 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Springer International Publishing |
record_format | MEDLINE/PubMed |
spelling | pubmed-78786572021-02-24 Effect of the Addition Frequency of 5-Azacytidine in Both Micro- and Macroscale Cultures Kadekar, Sandeep Barbe, Laurent Stoddart, Martin Varghese, Oommen P. Tenje, Maria Mestres, Gemma Cell Mol Bioeng Original Article INTRODUCTION: Human mesenchymal stem cells (hMSCs) have a great clinical potential for tissue regeneration purposes due to its multilineage capability. Previous studies have reported that a single addition of 5-azacytidine (5-AzaC) causes the differentiation of hMSCs towards a myocardial lineage. The aim of this work was to evaluate the effect of 5-AzaC addition frequency on hMSCs priming (i.e., indicating an early genetic differentiation) using two culture environments. METHODS: hMSCs were supplemented with 5-AzaC while cultured in well plates and in microfluidic chips. The impact of 5-AzaC concentration (10 and 20 μM) and addition frequency (once, daily or continuously), as well as of culture period (2 or 5 days) on the genetic upregulation of PPARγ (adipocytes), PAX3 (myoblasts), SOX9 (chondrocytes) and RUNX2 (osteoblasts) was evaluated. RESULTS: Daily delivering 5-AzaC caused a higher upregulation of PPARγ, SOX9 and RUNX2 in comparison to a single dose delivery, both under static well plates and dynamic microfluidic cultures. A particularly high gene expression of PPARγ (tenfold-change) could indicate priming of hMSCs towards adipocytes. CONCLUSIONS: Both macro- and microscale cultures provided results with similar trends, where addition frequency of 5-AzaC was a crucial factor to upregulate several genes. Microfluidics technology was proven to be a suitable platform for the continuous delivery of a drug and could be used for screening purposes in tissue engineering research. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s12195-020-00654-9) contains supplementary material, which is available to authorized users. Springer International Publishing 2020-10-06 /pmc/articles/PMC7878657/ /pubmed/33633814 http://dx.doi.org/10.1007/s12195-020-00654-9 Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Original Article Kadekar, Sandeep Barbe, Laurent Stoddart, Martin Varghese, Oommen P. Tenje, Maria Mestres, Gemma Effect of the Addition Frequency of 5-Azacytidine in Both Micro- and Macroscale Cultures |
title | Effect of the Addition Frequency of 5-Azacytidine in Both Micro- and Macroscale Cultures |
title_full | Effect of the Addition Frequency of 5-Azacytidine in Both Micro- and Macroscale Cultures |
title_fullStr | Effect of the Addition Frequency of 5-Azacytidine in Both Micro- and Macroscale Cultures |
title_full_unstemmed | Effect of the Addition Frequency of 5-Azacytidine in Both Micro- and Macroscale Cultures |
title_short | Effect of the Addition Frequency of 5-Azacytidine in Both Micro- and Macroscale Cultures |
title_sort | effect of the addition frequency of 5-azacytidine in both micro- and macroscale cultures |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7878657/ https://www.ncbi.nlm.nih.gov/pubmed/33633814 http://dx.doi.org/10.1007/s12195-020-00654-9 |
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