In Vitro Model for Hepatotoxicity Studies Based on Primary Human Hepatocyte Cultivation in a Perfused 3D Bioreactor System.
Accurate prediction of the potential hepatotoxic nature of new pharmaceuticals remains highly challenging. Therefore, novel in vitro models with improved external validity are needed to investigate hepatic metabolism and timely identify any toxicity of drugs in humans. In this study, we examined the effects of diclofenac, as a model substance with a known risk of hepatotoxicity in vivo, in a dynamic multi-compartment bioreactor using primary human liver cells. Biotransformation pathways of the drug and possible effects on metabolic activities, morphology and cell transcriptome were evaluated. Formation rates of diclofenac metabolites were relatively stable over the application period of seven days in bioreactors exposed to 300 µM diclofenac (300 µM bioreactors (300 µM BR)), while in bioreactors exposed to 1000 µM diclofenac (1000 µM BR) metabolite concentrations declined drastically. The biochemical data showed a significant decrease in lactate production and for the higher dose a significant increase in ammonia secretion, indicating a dose-dependent effect of diclofenac application. The microarray analyses performed revealed a stable hepatic phenotype of the cells over time and the observed transcriptional changes were in line with functional readouts of the system. In conclusion, the data highlight the suitability of the bioreactor technology for studying the hepatotoxicity of drugs in vitro.
Bioreactor Group, Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Charité-Universitätsmedizin Berlin, Campus Virchow-Klinikum, Berlin 13353, Germany. fanny.knoespel@charite.de.
Fanny
F
Knöspel
Janssen Research &amp; Development, Beerse 2340, Belgium. FJACOBS1@its.jnj.com.
Frank
F
Jacobs
Bioreactor Group, Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Charité-Universitätsmedizin Berlin, Campus Virchow-Klinikum, Berlin 13353, Germany. nora.freyer@charite.de.
Nora
N
Freyer
Department for General, Visceral and Transplantation Surgery, Charité-Universitätsmedizin Berlin, Campus Virchow-Klinikum, Berlin 13353, Germany. georg.damm@charite.de.
Georg
G
Damm
Janssen Research &amp; Development, Beerse 2340, Belgium. ADBONDT@its.jnj.com.
An
A
De Bondt
Janssen Research &amp; Development, Beerse 2340, Belgium. IVDWYNGA@its.jnj.com.
Ilse
I
van den Wyngaert
Janssen Research &amp; Development, Beerse 2340, Belgium. JSNOEYS@its.jnj.com.
Jan
J
Snoeys
Janssen Research &amp; Development, Beerse 2340, Belgium. MMONSHOU@ITS.JNJ.COM.
Mario
M
Monshouwer
Bioreactor Group, Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Charité-Universitätsmedizin Berlin, Campus Virchow-Klinikum, Berlin 13353, Germany. marco.richter@charite.de.
Marco
M
Richter
Bioreactor Group, Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Charité-Universitätsmedizin Berlin, Campus Virchow-Klinikum, Berlin 13353, Germany. nadja.strahl@charite.de.
Nadja
N
Strahl
Department for General, Visceral and Transplantation Surgery, Charité-Universitätsmedizin Berlin, Campus Virchow-Klinikum, Berlin 13353, Germany. daniel.seehofer@charite.de.
Daniel
D
Seehofer
Bioreactor Group, Berlin-Brandenburg Center for Regenerative Therapies (BCRT), Charité-Universitätsmedizin Berlin, Campus Virchow-Klinikum, Berlin 13353, Germany. katrin.zeilinger@charite.de.
Katrin
K
Zeilinger
D000894:Anti-Inflammatory Agents, Non-Steroidal / Q000378:metabolism / Q000633:toxicity*; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002478:Cells, Cultured; D004008:Diclofenac / Q000378:metabolism / Q000633:toxicity*; D004867:Equipment Design; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D018675:Toxicity Tests / Q000295:instrumentation*; D059467:Transcriptome
D000894:Anti-Inflammatory Agents, Non-Steroidal; D004008:Diclofenac
diclofenac; primary human hepatocytes, in vitro hepatotoxicity model; three-dimensional (3D) bioreactor