Relevance of the incubation period in cytotoxicity testing with primary human hepatocytes.

Fecha de publicación: 01/12/2018

Autores de IIS La Fe

Laia Tolosa Pardo

Autor
José Vicente Castell Ripoll

Autor

Participantes ajenos a IIS La Fe

Gu, XL
Albrecht, W
Edlund, K
Kappenberg, F
Rahnenfuhrer, J
Leist, M
Moritz, W
Godoy, P
Cadenas, C
Marchan, R
Brecklinghaus, T
Gardner, I
Han, B
Hengstler, JG
Stoeber, R

Grupos

Hepatología experimental y Trasplante hepático

Investigador Principal

Laia Tolosa Pardo
Unidad Mixta de Investigación en Hepatología Experimental

Investigador Principal

Ramiro Jover Atienza

Abstract

Primary human hepatocytes (PHHs) remain the gold standard for in vitro testing in the field of pharmacology and toxicology. One crucial parameter influencing the results of in vitro tests is the incubation period with test compounds. It has been suggested that longer incubation periods may be critical for the prediction of repeated dose toxicity. However, a study that systematically analyzes the relationship between incubation period and cytotoxicity in PHHs is not available. To close this gap, 30 compounds were tested in a concentration-dependent manner for cytotoxicity in cultivated cryopreserved PHHs (three donors per compound) for 1, 2 and 7days. The median of the EC 50 values of all compounds decreased 1.78-fold on day 2 compared to day 1, and 1.89-fold on day 7 compared to day 1. Median values of EC 50 ratios of all compounds at day 2 and day 7 were close to one but for individual compounds the ratio increased up to almost six. Strong correlations were obtained for EC 50 on day 1 and day 7 (R = 0.985; 95% CI 0.960-0.994), day 1 and day 2 (R = 0.964; 95% CI 0.910-0.986), as well as day 2 and day 7 (R = 0.981; 95% CI 0.955-0.992). However, compound specific differences also occurred. Whereas, for example, busulfan showed a relatively strong increase on day 7 compared to day 1, cytotoxicity of acetaminophen did not increase during longer incubation periods. To validate the observed correlations, a publicly available data set, containing data on the cytotoxicity of human hepatocytes cultivated as spheroids for incubation periods of 5 and 14days, was analyzed. A high correlation coefficient of EC 50 values at day 5 and day 14 was obtained (R = 0.894; 95% CI 0.798-0.945). In conclusion, the median cytotoxicity of the test compounds increased between 1 and 2days of incubation, with no or only a minimal further increase until day 7. It remains to be studied whether the different results obtained for some individual compounds after longer exposure periods would correspond better to human-repeated dose toxicity.

Datos de la publicación

ISSN/ISSNe:: 0340-5761, 1432-0738
Tipo:: Article
Páginas:: 3505-3515
DOI:: 10.1007/s00204-018-2302-0
Factor de Impacto:: 1,609 SCImago ℠
Cuartil:: Q1 SCImago ℠

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Métricas

Filiaciones

Gu, XL:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), 44139, Dortmund, Germany

Tech Univ Dortmund IfADo, Leibniz Res Ctr Working Environm & Human Factors, D-44139 Dortmund, Germany
Albrecht, W:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), 44139, Dortmund, Germany

Tech Univ Dortmund IfADo, Leibniz Res Ctr Working Environm & Human Factors, D-44139 Dortmund, Germany
Edlund, K:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), 44139, Dortmund, Germany

Tech Univ Dortmund IfADo, Leibniz Res Ctr Working Environm & Human Factors, D-44139 Dortmund, Germany
Kappenberg, F:: Department of Statistics, TU Dortmund University, 44227, Dortmund, Germany

TU Dortmund Univ, Dept Stat, D-44227 Dortmund, Germany
Rahnenfuhrer, J:: Department of Statistics, TU Dortmund University, 44227, Dortmund, Germany

TU Dortmund Univ, Dept Stat, D-44227 Dortmund, Germany
Leist, M:: In Vitro Toxicology and Biomedicine, Department of Biology, University of Konstanz, Universitätsstr. 10, PO Box M657, 78457, Konstanz, Germany

Univ Konstanz, Dept Biol, In Vitro Toxicol & Biomed, Univ Str 10,POB M657, D-78457 Constance, Germany
Moritz, W:: InSphero AG, Technoparkstr. 1, 8005, Zürich, Switzerland

InSphero AG, Technopk Str 1, CH-8005 Zurich, Switzerland
Godoy, P:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), 44139, Dortmund, Germany

Tech Univ Dortmund IfADo, Leibniz Res Ctr Working Environm & Human Factors, D-44139 Dortmund, Germany
Cadenas, C:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), 44139, Dortmund, Germany

Tech Univ Dortmund IfADo, Leibniz Res Ctr Working Environm & Human Factors, D-44139 Dortmund, Germany
Marchan, R:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), 44139, Dortmund, Germany

Tech Univ Dortmund IfADo, Leibniz Res Ctr Working Environm & Human Factors, D-44139 Dortmund, Germany
Brecklinghaus, T:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), 44139, Dortmund, Germany

Tech Univ Dortmund IfADo, Leibniz Res Ctr Working Environm & Human Factors, D-44139 Dortmund, Germany
Pardo, LT:: Instituto de Investigación. Unit for Cell Therapy, University La Fe Hospital, Valencia, Spain

Univ La Fe Hosp, Unit Cell Therapy, Valencia, Spain
Castell, JV:: Instituto de Investigación. Unit for Cell Therapy, University La Fe Hospital, Valencia, Spain

Univ La Fe Hosp, Unit Cell Therapy, Valencia, Spain
Gardner, I:: Simcyp, Sheffield, UK

Simcyp, Sheffield, S Yorkshire, England
Hengstler, JG:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), 44139, Dortmund, Germany.

Tech Univ Dortmund IfADo, Leibniz Res Ctr Working Environm & Human Factors, D-44139 Dortmund, Germany
Stoeber, R:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), 44139, Dortmund, Germany

Tech Univ Dortmund IfADo, Leibniz Res Ctr Working Environm & Human Factors, D-44139 Dortmund, Germany

Keywords

Cell-titer-blue, Hepatotoxicity, Incubation period, Primary human hepatocyte

Proyectos asociados

DESARROLLO DE UN MODELO HEPATOCELULAR HUMANO DIFERENCIADO PARA ESTUDIOS DE METABOLISMO Y POTENCIAL INDUCTOR DE NUEVOS FARMACOS

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A-CUTE-TOX

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LSHB-CT-2004-512051-ACUTE TOX . FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2005

DESARROLLO DE NUEVAS ESTRATEGIAS DIAGNOSTICAS BASADAS EN EL ESTUDIO DE LA CAPACIDAD METABOLIZANTE DE LA PIEL Y DEL PAPEL DE LAS CELULAS DENDITRICAS EN LA INDUCCION DE LAS REACCIONES ALERGICAS FRENTE A FARMACOS

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

PARTIC PI05/2290 - FUND IMABIS . INSTITUTO DE SALUD CARLOS III; FUNDACION FIMABIS . 2007

DEVELOPMENT OF A HIGH THROUGHPUT GENOMICS-BASED TEST FOR ASSESSING GENOTOXIC AND CARCINOGENIC PROPERTIES OF CHEMICAL COMPOUNDS IN VITRO

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CARCINOGENOMICS . COMISION EUROPEA . 2006

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LSHB-CT-2006-037499 - LINTOP . FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2005

MECANISMOS TRNACRIPCIONALES IMPLICADOS EN LA ETIOLOGIA DEL HIGADO GRASO NO ALCOHOLICO. ESTUDIOS EN UN MODELO CELULAR HUMANO DE ESTEATOSIS Y APLICACION AL TRANSPLANTE DE HEPATOCITOS EN TERAPIA CELULAR.

Investigador Principal: RAMIRO JOVER ATIENZA

PI07/0550 . INSTITUTO DE SALUD CARLOS III . 2007

TU HOSPITAL INVESTIGA PARA TI

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CCT005-07-00936 . 2007

PROYECTO CAIBER

CAI08/01/0061 . INSTITUTO DE SALUD CARLOS III; FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2009

FORINVITOX

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RED DE INNOVACION EN TECNOLOGIAS MEDICAS Y SANITARIAS - RED ITEMAS

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RD09/0077/00156 . INSTITUTO DE SALUD CARLOS III . 2010

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Investigador Principal: RAMIRO JOVER ATIENZA

PI10/00194 . INSTITUTO DE SALUD CARLOS III . 2010

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Investigador Principal: MARÍA TERESA DONATO MARTÍN

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