EXCLI J EXCLI Journal 1611-2156 Leibniz Research Centre for Working Environment and Human Factors 2018-2017 10.17179/excli2018-2017 Doc1180 Editorial material Highlight report: Cell type selection for toxicity testing Bolt H. M. * 1 IfADo - Leibniz Research Centre for Working Environment and Human Factors, Dortmund, GERMANY *To whom correspondence should be addressed: H. M. Bolt, IfADo - Leibniz Research Centre for Working Environment and Human Factors, Dortmund, GERMANY, E-mail: bolt@ifado.de 18 12 2018 2018 17 1180 1181 17 12 2018 18 12 2018 Copyright © 2018 Bolt 2018

This is an Open Access article distributed under the terms of the Creative Commons Attribution Licence (http://creativecommons.org/licenses/by/4.0/) You are free to copy, distribute and transmit the work, provided the original author and source are credited.

This article is available from http://www.excli.de/vol17/Bolt_18122018_proof.pdf


An important step in in vitro test system development is the choice of an adequate cell line which depends on the intended application of the assay. In a recent study, Tuuli Karhu and colleagues from Helsinki University compared a set of cell lines for their susceptibility towards eight GATA4 targeting compounds (Karhu et al., 2018[7]). GATA4 is a transcription factor involved in cardiac development (Gupta et al., 2013[6]; Kikuchi et al., 2010[8]; Rysä et al., 2010[15]; Pikkarainen et al., 2004[13]). The goal of the study was to identify which cell line allows the most sensitive cytotoxicity screening of these compounds. The tested cell lines included the myoblast cell line H9c2 established from rat myocardium; primary neonatal rat cardiac fibroblasts; mouse embryonic fibroblasts; mouse embryonic stem cells (mECSs), mouse embryonic stem cell derivatives from day 5 embryoid bodies; induced pluripotent human stem cells (hiPSC); and hiPSC-derived cardiomyocytes. The most susceptible cell lines towards the set of test compounds were hiPSC and mESC, while cardiomyocytes, fibroblasts and H9c2 cells were most resistant (Karhu et al., 2018[7]). Of course screening for the most sensitive cell line does not guarantee that the test cells will be most relevant for the human in vivo situation. However, if one is interested in a cytotoxicity screening system with the highest sensitivity, the recommendation of the authors to further use hiPSC seems reasonable.

In recent years, the development of stem cell based test systems has been a major focus of research (Leist et al., 2017[10]; Godoy et al., 2013[3]; Krug et al., 2013[9]). The most frequently applied strategy is to expose stem cells to test compounds, when they differentiate to more mature cell types (Shinde et al., 2017[17]; Pallocca et al., 2016[12]).

This approach has been used for developmental neurotoxicity (Waldmann et al., 2014[18]; Meganathan et al., 2015[11]; Weng et al., 2014[19]; Rempel et al., 2015[14]) and for cardiotoxicity (Chaudhari et al., 2016[1][2]; Sampaio et al., 2016[16]) testing. While tests that analyze the influence of compounds on the differentiation process are already successfully applied, it still remains a challenge to generate mature cell types, e.g. hepatocytes that closely resemble the primary cells in an adult organ (Godoy et al., 2016[5], 2018[4]). Although much progress has been achieved in stem cell based test system development, systematic analysis of human in vivo relevance still remains a major challenge.

Chaudhari U Nemade H Gaspar JA Hescheler J Hengstler JG Sachinidis A MicroRNAs as early toxicity signatures of doxorubicin in human-induced pluripotent stem cell-derived cardiomyocytes Arch Toxicol 2016 90 3087 3098 Chaudhari U Nemade H Wagh V Gaspar JA Ellis JK Srinivasan SP Identification of genomic biomarkers for anthracycline-induced cardiotoxicity in human iPSC-derived cardiomyocytes: an in vitro repeated exposure toxicity approach for safety assessment Arch Toxicol 2016 90 2763 2777 Godoy P Hewitt NJ Albrecht U Andersen ME Ansari N Bhattacharya S Recent advances in 2D and 3D in vitro systems using primary hepatocytes, alternative hepatocyte sources and non-parenchymal liver cells and their use in investigating mechanisms of hepatotoxicity, cell signaling and ADME Arch Toxicol 2013 87 1315 1530 Godoy P Schmidt-Heck W Hellwig B Nell P Feuerborn D Rahnenführer J Assessment of stem cell differentiation based on genome-wide expression profiles Philos Trans R Soc Lond B Biol Sci 2018 373 1750 Godoy P Widera A Schmidt-Heck W Campos G Meyer C Cadenas C Gene network activity in cultivated primary hepatocytes is highly similar to diseased mammalian liver tissue Arch Toxicol 2016 90 2513 2529 Gupta V Gemberling M Karra R Rosenfeld GE Evans T Poss KD An injury-responsive gata4 program shapes the zebrafish cardiac ventricle Curr Biol 2013 23 1221 1227 Karhu ST Välimäki MJ Jumppanen M Kinnunen SM Pohjolainen L Leigh RS Stem cells are the most sensitive screening tool to identify toxicity of GATA4-targeted novel small-molecule compounds Arch Toxicol 9 Jul 2018 epub ahead of print 10.1007/s00204-018-2257-1. Available from: http://dx.doi.org/10.1007/s00204-018-2257-1 Kikuchi K Holdway JE Werdich AA Anderson RM Fang Y Egnaczyk GF Primary contribution to zebrafish heart regeneration by gata4(+) cardiomyocytes Nature 2010 464 7288 601 605 Krug AK Kolde R Gaspar JA Rempel E Balmer NV Meganathan K Human embryonic stem cell-derived test systems for developmental neurotoxicity: a transcriptomics approach Arch Toxicol 2013 87 123 143 Leist M Ghallab A Graepel R Marchan R Hassan R Bennekou SH Adverse outcome pathways: opportunities, limitations and open questions Arch Toxicol 2017 91 3477 3505 Meganathan K Jagtap S Srinivasan SP Wagh V Hescheler J Hengstler J Neuronal developmental gene and miRNA signatures induced by histone deacetylase inhibitors in human embryonic stem cells Cell Death Dis 2015 6 e1756 Pallocca G Grinberg M Henry M Frickey T Hengstler JG Waldmann T Identification of transcriptome signatures and biomarkers specific for potential developmental toxicants inhibiting human neural crest cell migration Arch Toxicol 2016 90 159 180 Pikkarainen S Tokola H Kerkelä R Ruskoaho H GATA transcription factors in the developing and adult heart Cardiovasc Res 2004 63 196 207 Rempel E Hoelting L Waldmann T Balmer NV Schildknecht S Grinberg M A transcriptome-based classifier to identify developmental toxicants by stem cell testing: design, validation and optimization for histone deacetylase inhibitors Arch Toxicol 2015 89 1599 1618 Rysä J Tenhunen O Serpi R Soini Y Nemer M Leskinen H GATA-4 is an angiogenic survival factor of the infarcted heart Circ Heart Fail 2010 3 440 450 Sampaio SF Branco AF Wojtala A Vega-Naredo I Wieckowski MR Oliveira PJ p66Shc signaling is involved in stress responses elicited by anthracycline treatment of rat cardiomyoblasts Arch Toxicol 2016 90 1669 1684 Shinde V Hoelting L Srinivasan SP Meisig J Meganathan K Jagtap S Definition of transcriptome-based indices for quantitative characterization of chemically disturbed stem cell development: introduction of the STOP-Toxukn and STOP-Toxukk tests Arch Toxicol 2017 91 839 864 Waldmann T Rempel E Balmer NV König A Kolde R Gaspar JA Design principles of concentration-dependent transcriptome deviations in drug-exposed differentiating stem cells Chem Res Toxicol 2014 27 408 420 Weng MK Natarajan K Scholz D Ivanova VN Sachinidis A Hengstler JG Lineage-specific regulation of epigenetic modifier genes in human liver and brain PLoS One 2014 9 7 e102035