Effect of human patient plasma ex vivo treatment on gene expression and progenitor cell activation of primary human liver cells in multi‐compartment 3D perfusion bioreactors for extra‐corporeal liver support

Schmelzer, Eva; Mutig, Kerim; Schrade, Petra; Bachmann, S.; Gerlach, Jörg C.; Zeilinger, Katrin

doi:10.1002/bit.22283

Cited by 46 publications

(41 citation statements)

References 53 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…1C). A detailed description of the bioreactor structure is given elsewhere (Schmelzer et al, 2009). In this study, a downscaled bioreactor with a cell compartment volume of 2 ml was used (Fig.…”

Section: Methodsmentioning

confidence: 99%

“…It has been shown that porcine or human liver cells retain in vivo-like properties and are arranged in tissue-like structures including formation of biliary canalicular networks and neo-sinusoids when cultured in a perfused 3D bioreactor (Zeilinger et al, 2004;Schmelzer et al, 2009). More importantly, liver-specific functions such as urea and albumin synthesis, glucose metabolism, and P450 activities were all maintained for at least 14 days in this type of bioreactor (Zeilinger et al, 2002).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Cytochrome P450-Dependent Metabolism in HepaRG Cells Cultured in a Dynamic Three-Dimensional Bioreactor

Darnell¹,

Schreiter²,

Zeilinger³

et al. 2011

Drug Metab Dispos

Self Cite

View full text Add to dashboard Cite

ABSTRACT:Reliable and stable in vitro cellular systems maintaining specific liver functions important for drug metabolism and disposition are urgently needed in preclinical drug discovery and development research. The cell line HepaRG exhibits promising properties such as expression and function of drug-metabolizing enzymes and transporter proteins, which resemble those found in freshly isolated human hepatocytes. In this study, HepaRG cells were cultured up to 68 days in a three-dimensional multicompartment capillary membrane bioreactor, which enables high-density cell culture under dynamic conditions. The activity of drug-metabolizing cytochrome P450 (P450) enzymes was investigated by a cocktail of substrates for CYP1A1/2 (phenacetin), CYP2C9 (diclofenac), CYP2B6 (bupropion), and CYP3A4 (midazolam). The model P450 substrates, which were introduced to the bioreactor system mimicking in vivo bolus doses, showed stable metabolism over the entire experimental period of several weeks with the exception of bupropion hydroxylase, which increased over time. Ketoconazole treatment decreased the CYP3A4 activity by 69%, and rifampicin induced the CYP3A4-and CYP2B6-dependent activity 6-fold, which predicts well the magnitude of changes observed in vivo. Moreover, polarity of transporter expression and formation of tissue-like structures including bile canaliculi were demonstrated by immune histochemistry. The long-lasting bioreactor system using HepaRG cells thus provides a promising and stable liver-like in vitro model for continuous investigations of the hepatic kinetics of drugs and of drug-drug interactions, which well predict the situation in vivo in humans.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Cytochrome P450-Dependent Metabolism in HepaRG Cells Cultured in a Dynamic Three-Dimensional Bioreactor

Darnell¹,

Schreiter²,

Zeilinger³

et al. 2011

Drug Metab Dispos

Self Cite

View full text Add to dashboard Cite

show abstract

“…To mimic the situation in the liver, fresh human hepatocytes can be cultured in bioreactors, which enables a 3D structure, cell-cell contact, and a constant medium flow and oxygen supply that are important for the intracellular functions and maintenance of the cell polarity (Tilles et al, 2001;Zeilinger et al, 2004;Schmelzer et al, 2009;Vinci et al, 2011). It has been shown previously that fresh human hepatocytes can maintain their liver-specific functions such as urea and albumin synthesis, glucose metabolism, and P450 activities for at least 14 days in 3D cultures (Zeilinger et al, 2002.…”

Section: Introductionmentioning

confidence: 99%

In Vitro Evaluation of Major In Vivo Drug Metabolic Pathways Using Primary Human Hepatocytes and HepaRG Cells in Suspension and a Dynamic Three-Dimensional Bioreactor System

Darnell

Ulvestad

Ellis

et al. 2012

J Pharmacol Exp Ther

View full text Add to dashboard Cite

Major human specific metabolites, not detected during in vivo and in vitro preclinical studies, may cause unexpected drug interactions and toxicity in human and delays in clinical programs. Thus, reliable preclinical tools for the detection of major human metabolites are of high importance. The aim of this study was to compare major drug metabolic pathways in HepaRG cells, a human hepatoma cell line, to fresh human hepatocytes, cryopreserved human hepatocytes, and human in vivo data. Furthermore, the maintenance of cytochrome P450 (P450) and UDP-glucuronosyltransferase (UGT) activities in a dynamic three-dimensional (3D) bioreactor were evaluated over time by using HepaRG cells and human hepatocytes.14 Cdiclofenac and a candidate from AstraZeneca's drug development program, 14 C-AZD6610, which are metabolized by P450 and UGT in vivo, were used as model substrates. The proportion of relevant biotransformation pathways of the investigated drug was clearly different in the various cell systems. The hydroxylation route was favored in primary human hepatocytes, whereas the glucuronidation route was favored in HepaRG cells. The human in vivo metabolite profile of AZD6610 was best represented by human hepatocytes, whereas all major diclofenac metabolites were detected in HepaRG cells. Moreover, the metabolite profiles in cryopreserved and fresh human hepatocytes were essentially the same. The liver bioreactor using both fresh human hepatocytes and HepaRG cells retained biotransformation capacity over 1 week. Thus, the incubation time can be increased from a few hours in suspension to several days in 3D cultures, which opens up for detection of metabolites from slowly metabolized drugs.

show abstract

“…As well as being used clinically [21], they are being utilised as in vitro models, like the scaling down of a 3D perfusion multicompartment hollow fibre liver bioreactor for use in in vitro pharmacological studies [22]. This bioreactor consists of three interwoven sets of hollow fibre, two for counter current medium perfusion and one for gas supply (Fig 1).…”

Section: Status Of Bioreactors In the Development Of In Vitro Modelsmentioning

confidence: 99%

The use of bioreactors as in vitro models in pharmaceutical research

Ginai

Elsby

Hewitt

et al. 2013

Drug Discovery Today

View full text Add to dashboard Cite

Word TeaserThe development of dynamic 3D bioreactor based systems as in vitro models for use in DMPK studies. Author BiographiesMaaria Ginai graduated with a first Class Bachelors degree with honours in Biomedical Science from the University of Kent (UK) in 2010. She is currently studying a BBSRC CASEfunded PhD at the Centre for Biological Engineering at Loughborough University in the area of bioartificial device progression to in vitro models for use in the pharmaceutical industry. This project is supported by AstraZeneca.Christopher J. Hewitt has a first Class Bachelors degree in Biology from Royal Holloway College, University of London (UK) and a PhD in Chemical Engineering from the University of Birmingham (UK). He is Director of the £7.3M EPSRC Doctoral Training Centre in Regenerative Medicine and co-founder of the £2M Centre for Biological Engineering at Loughborough University (UK). He also leads the Cell Technologies research group whose work spans the Engineering/Life science interface seeking to understand the interaction of the organism with the engineering environment within such diverse areas as microbial fermentation, bio-transformation, cell culture and mostly recently regenerative medicine bioprocessing.Karen Coopman has a first Class Bachelors degree in Pharmacology from the University of Bristol (UK) and a PhD from the Department of Pharmacy and Pharmacology at the University of Bath (UK). She was appointed to a lectureship at Loughborough University, where she is the Operations Manager of the EPSRC-funded Doctoral Training Centre in Regenerative Medicine. She co-leads the Cell Technologies research group within University's Centre for Biological Engineering and is currently a member of the EPSRC's Early Career Forum in Manufacturing Research. The overarching themes of Karen's research are the manufacture of cellular therapies and the use of cells in the drug discovery process. AbstractBringing a new drug to market is costly in terms of capital and time investments, and any development issues encountered in late stage clinical trials may often be due to in vitro -in vivo extrapolations (IVIVE) not accurately reflecting clinical outcome. In the discipline of drug metabolism and pharmacokinetics (DMPK), current in vitro cellular methods do not provide the 3D structure and function of organs found in vivo, so new dynamic methods need to be established to aid improvement of IVIVE. This review will highlight the importance of model progression into dynamic systems for use within drug development, focussing on devices developed currently in the areas of the liver and blood-brain barrier, and the potential to develop models for other organ systems such as the kidney.

show abstract

Effect of human patient plasma ex vivo treatment on gene expression and progenitor cell activation of primary human liver cells in multi‐compartment 3D perfusion bioreactors for extra‐corporeal liver support

Cited by 46 publications

References 53 publications

Cytochrome P450-Dependent Metabolism in HepaRG Cells Cultured in a Dynamic Three-Dimensional Bioreactor

Cytochrome P450-Dependent Metabolism in HepaRG Cells Cultured in a Dynamic Three-Dimensional Bioreactor

In Vitro Evaluation of Major In Vivo Drug Metabolic Pathways Using Primary Human Hepatocytes and HepaRG Cells in Suspension and a Dynamic Three-Dimensional Bioreactor System

The use of bioreactors as in vitro models in pharmaceutical research

Contact Info

Product

Resources

About