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The effect of a number of H9C2 rat cardiomyocytes passage on repeatability of cytotoxicity study results

The embryonic cardiomyocyte cell line H9C2 is commonly used in numerous in vitro studies, including cardiotoxicity analyses of new drugs. So far no results were published for studies on cell parameters variability during the cell line ageing process. For this reason the aim of the study was to evalu...

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Autores principales: Witek, Piotr, Korga, Agnieszka, Burdan, Franciszek, Ostrowska, Marta, Nosowska, Beata, Iwan, Magdalena, Dudka, Jarosław
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Netherlands 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5101311/
https://www.ncbi.nlm.nih.gov/pubmed/26946144
http://dx.doi.org/10.1007/s10616-016-9957-2
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author Witek, Piotr
Korga, Agnieszka
Burdan, Franciszek
Ostrowska, Marta
Nosowska, Beata
Iwan, Magdalena
Dudka, Jarosław
author_facet Witek, Piotr
Korga, Agnieszka
Burdan, Franciszek
Ostrowska, Marta
Nosowska, Beata
Iwan, Magdalena
Dudka, Jarosław
author_sort Witek, Piotr
collection PubMed
description The embryonic cardiomyocyte cell line H9C2 is commonly used in numerous in vitro studies, including cardiotoxicity analyses of new drugs. So far no results were published for studies on cell parameters variability during the cell line ageing process. For this reason the aim of the study was to evaluate the effect of a number of H9C2 rat embryonic cardiomyocytes passages on repeatability of study results for selected cytotoxicity parameters, with doxorubicin as a model toxic agent. The cultures were passaged twenty-five times. Cells from passage 1, 5, 10, 15, 20 and 25 were treated with doxorubicin for 24 h. Then drug cytotoxicity was evaluated with the MTT test and additionally the nuclear factor erythroid 2–related factor (Nrf2) gene expression was examined. The analysis of oxidative stress intensity and cell morphology was also assessed. The microscopic appearance of cells indicates that untreated cardiomyocytes morphology changes as well as sensitivity to toxic effects increases with the number of passages. Also an increase in oxidative stress in cells occurs with further passaging of cardiomyocytes. Statistical significance of differences in conducted tests results depended on doxorubicin concentration but in many cases the H9C2 line was found to be a reliable in vitro model only for the first five passages. For this reason it is important to take into consideration that further culturing of cardiomyocytes may not ensure repeatability of study results due to the culture ageing.
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spelling pubmed-51013112016-11-21 The effect of a number of H9C2 rat cardiomyocytes passage on repeatability of cytotoxicity study results Witek, Piotr Korga, Agnieszka Burdan, Franciszek Ostrowska, Marta Nosowska, Beata Iwan, Magdalena Dudka, Jarosław Cytotechnology Original Article The embryonic cardiomyocyte cell line H9C2 is commonly used in numerous in vitro studies, including cardiotoxicity analyses of new drugs. So far no results were published for studies on cell parameters variability during the cell line ageing process. For this reason the aim of the study was to evaluate the effect of a number of H9C2 rat embryonic cardiomyocytes passages on repeatability of study results for selected cytotoxicity parameters, with doxorubicin as a model toxic agent. The cultures were passaged twenty-five times. Cells from passage 1, 5, 10, 15, 20 and 25 were treated with doxorubicin for 24 h. Then drug cytotoxicity was evaluated with the MTT test and additionally the nuclear factor erythroid 2–related factor (Nrf2) gene expression was examined. The analysis of oxidative stress intensity and cell morphology was also assessed. The microscopic appearance of cells indicates that untreated cardiomyocytes morphology changes as well as sensitivity to toxic effects increases with the number of passages. Also an increase in oxidative stress in cells occurs with further passaging of cardiomyocytes. Statistical significance of differences in conducted tests results depended on doxorubicin concentration but in many cases the H9C2 line was found to be a reliable in vitro model only for the first five passages. For this reason it is important to take into consideration that further culturing of cardiomyocytes may not ensure repeatability of study results due to the culture ageing. Springer Netherlands 2016-03-05 2016-12 /pmc/articles/PMC5101311/ /pubmed/26946144 http://dx.doi.org/10.1007/s10616-016-9957-2 Text en © The Author(s) 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Original Article
Witek, Piotr
Korga, Agnieszka
Burdan, Franciszek
Ostrowska, Marta
Nosowska, Beata
Iwan, Magdalena
Dudka, Jarosław
The effect of a number of H9C2 rat cardiomyocytes passage on repeatability of cytotoxicity study results
title The effect of a number of H9C2 rat cardiomyocytes passage on repeatability of cytotoxicity study results
title_full The effect of a number of H9C2 rat cardiomyocytes passage on repeatability of cytotoxicity study results
title_fullStr The effect of a number of H9C2 rat cardiomyocytes passage on repeatability of cytotoxicity study results
title_full_unstemmed The effect of a number of H9C2 rat cardiomyocytes passage on repeatability of cytotoxicity study results
title_short The effect of a number of H9C2 rat cardiomyocytes passage on repeatability of cytotoxicity study results
title_sort effect of a number of h9c2 rat cardiomyocytes passage on repeatability of cytotoxicity study results
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5101311/
https://www.ncbi.nlm.nih.gov/pubmed/26946144
http://dx.doi.org/10.1007/s10616-016-9957-2
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