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Genotoxic Effects of Culture Media on Human Pluripotent Stem Cells

Culture conditions play an important role in regulating the genomic integrity of Human Pluripotent Stem Cells (HPSCs). We report that HPSCs cultured in Essential 8 (E8) and mTeSR, two widely used media for feeder-free culturing of HPSCs, had many fold higher levels of ROS and higher mitochondrial po...

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Autores principales: Prakash Bangalore, Megha, Adhikarla, Syama, Mukherjee, Odity, Panicker, Mitradas M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5297241/
https://www.ncbi.nlm.nih.gov/pubmed/28176872
http://dx.doi.org/10.1038/srep42222
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author Prakash Bangalore, Megha
Adhikarla, Syama
Mukherjee, Odity
Panicker, Mitradas M.
author_facet Prakash Bangalore, Megha
Adhikarla, Syama
Mukherjee, Odity
Panicker, Mitradas M.
author_sort Prakash Bangalore, Megha
collection PubMed
description Culture conditions play an important role in regulating the genomic integrity of Human Pluripotent Stem Cells (HPSCs). We report that HPSCs cultured in Essential 8 (E8) and mTeSR, two widely used media for feeder-free culturing of HPSCs, had many fold higher levels of ROS and higher mitochondrial potential than cells cultured in Knockout Serum Replacement containing media (KSR). HPSCs also exhibited increased levels of 8-hydroxyguanosine, phospho-histone-H2a.X and p53, as well as increased sensitivity to γ-irradiation in these two media. HPSCs in E8 and mTeSR had increased incidence of changes in their DNA sequence, indicating genotoxic stress, in addition to changes in nucleolar morphology and number. Addition of antioxidants to E8 and mTeSR provided only partial rescue. Our results suggest that it is essential to determine cellular ROS levels in addition to currently used criteria i.e. pluripotency markers, differentiation into all three germ layers and normal karyotype through multiple passages, in designing culture media.
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spelling pubmed-52972412017-02-13 Genotoxic Effects of Culture Media on Human Pluripotent Stem Cells Prakash Bangalore, Megha Adhikarla, Syama Mukherjee, Odity Panicker, Mitradas M. Sci Rep Article Culture conditions play an important role in regulating the genomic integrity of Human Pluripotent Stem Cells (HPSCs). We report that HPSCs cultured in Essential 8 (E8) and mTeSR, two widely used media for feeder-free culturing of HPSCs, had many fold higher levels of ROS and higher mitochondrial potential than cells cultured in Knockout Serum Replacement containing media (KSR). HPSCs also exhibited increased levels of 8-hydroxyguanosine, phospho-histone-H2a.X and p53, as well as increased sensitivity to γ-irradiation in these two media. HPSCs in E8 and mTeSR had increased incidence of changes in their DNA sequence, indicating genotoxic stress, in addition to changes in nucleolar morphology and number. Addition of antioxidants to E8 and mTeSR provided only partial rescue. Our results suggest that it is essential to determine cellular ROS levels in addition to currently used criteria i.e. pluripotency markers, differentiation into all three germ layers and normal karyotype through multiple passages, in designing culture media. Nature Publishing Group 2017-02-08 /pmc/articles/PMC5297241/ /pubmed/28176872 http://dx.doi.org/10.1038/srep42222 Text en Copyright © 2017, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Prakash Bangalore, Megha
Adhikarla, Syama
Mukherjee, Odity
Panicker, Mitradas M.
Genotoxic Effects of Culture Media on Human Pluripotent Stem Cells
title Genotoxic Effects of Culture Media on Human Pluripotent Stem Cells
title_full Genotoxic Effects of Culture Media on Human Pluripotent Stem Cells
title_fullStr Genotoxic Effects of Culture Media on Human Pluripotent Stem Cells
title_full_unstemmed Genotoxic Effects of Culture Media on Human Pluripotent Stem Cells
title_short Genotoxic Effects of Culture Media on Human Pluripotent Stem Cells
title_sort genotoxic effects of culture media on human pluripotent stem cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5297241/
https://www.ncbi.nlm.nih.gov/pubmed/28176872
http://dx.doi.org/10.1038/srep42222
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