Differentiation-Promoting Medium Additives for Hepatocyte Cultivation and Cryopreservation

Gouliarmou, Varvara; Pelkonen, Olavi; Coecke, Sandra

doi:10.1007/978-1-4939-2074-7_10

Cited by 5 publications

(3 citation statements)

References 94 publications

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“…For long‐term culturing, various strategies have been used to maintain the differentiation status of the isolated hepatocytes, but the main focus is still on the strategy that employs differentiation‐promoting compounds in the culture medium. An overview on various antidifferentiation procedures and cryopreservation agents has been published (Gouliarmou et al, 2015 ). Metabolically competent cryopreserved human primary hepatocyte culture used for the validation of CYP induction has been recently published (Bernasconi et al, 2019 ) and protocols are available (TSAR, TM2009‐13), where also most important primary literature sources have been referenced.…”

Section: Appendix C – Experimental Test Conditionsmentioning

confidence: 99%

Scientific Opinion of the Scientific Panel on Plant Protection Products and their Residues (PPR Panel) on testing and interpretation of comparative in vitro metabolism studies

Hernández‐Jerez¹,

Adriaanse²,

Aldrich³

et al. 2021

EFS2

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EFSA asked the Panel on Plant Protection Products and their residues to deliver a Scientific Opinion on testing and interpretation of comparative in vitro metabolism studies for both new active substances and existing ones. The main aim of comparative in vitro metabolism studies of pesticide active substances is to evaluate whether all significant metabolites formed in the human in vitro test system, as a surrogate of the in vivo situation, are also present at comparable level in animal species tested in toxicological studies and, therefore, if their potential toxicity has been appropriately covered by animal studies. The studies may also help to decide which animal model, with regard to a particular compound, is the most relevant for humans. In the experimental strategy, primary hepatocytes in suspension or culture are recommended since hepatocytes are considered the most representative in vitro system for prediction of in vivo metabolites. The experimental design of 3 9 3 9 3 (concentrations, time points, technical replicates, on pooled hepatocytes) will maximise the chance to identify unique (UHM) and disproportionate (DHM) human metabolites. When DHM and UHM are being assessed, test item-related radioactivity recovery and metabolite profile are the most important parameters. Subsequently, structural characterisation of the assigned metabolites is performed with appropriate analytical techniques. In toxicological assessment of metabolites, the uncertainty factor approach is the first alternative to testing option, followed by new approach methodologies (QSAR, read-across, in vitro methods), and only if these fail, in vivo animal toxicity studies may be performed. Knowledge of in vitro metabolites in human and animal hepatocytes would enable toxicological evaluation of all metabolites of concern, and, furthermore, add useful pieces of information for detection and evaluation of metabolites in different matrices (crops, livestock, environment), improve biomonitoring efforts via better toxicokinetic understanding, and ultimately, develop regulatory schemes employing physiologically based or physiology-mimicking in silico and/or in vitro test systems to anticipate the exposure of humans to potentially hazardous substances in plant protection products.

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Section: Appendix C – Experimental Test Conditionsmentioning

confidence: 99%

Scientific Opinion of the Scientific Panel on Plant Protection Products and their Residues (PPR Panel) on testing and interpretation of comparative in vitro metabolism studies

Hernández‐Jerez¹,

Adriaanse²,

Aldrich³

et al. 2021

EFS2

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“…A common method of storing cells including hepatocytes for a longer time is the cryopreservation by freezing cells in cryoprotective agents below −130°C (8)(9)(10). Cryoprotective agents prevent the formation of intracellular ice crystals and reduce the osmotic stress caused by cellular dehydration during the freezing process (10,11).…”

Section: Introductionmentioning

confidence: 99%

“…This solution was developed in the 1980s for clinical application ( 16 ). As it allows a proper preservation of human liver grafts, the University of Wisconsin solution often serves as freezing medium for the cryopreservation of hepatocytes ( 8 , 15 ).…”

Section: Introductionmentioning

confidence: 99%

Comparative cryopreservation of bovine and porcine primary hepatocytes

et al. 2023

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The isolation of primary hepatocytes from liver tissue of farm animals yields a very high number of cells, and a part of them can be stored by cryopreservation for future experiments. As no experience exists with the cryopreservation of hepatocytes from cattle, our study aimed at the cryopreservation of bovine hepatocytes by use of different protocols compared with the cryopreservation of hepatocytes from pig. We tested different freezing media (William’s Medium E vs. University of Wisconsin solution), cryoprotectants (dimethyl sulfoxide with vs. without trehalose as additional additive), freezing systems (standard freezing container vs. controlled-rate freezer) and freezing times (4 vs. 28 d). These tests identified a general influence of species and freezing systems, whereas the influence of freezing media, trehalose additive and freezing time was less or not obvious. In this regard, we determined a mean recovery of 30% of bovine hepatocytes and 55% of porcine hepatocytes cryopreserved in a controlled-rate freezer, whereas the rates were about 10% less when hepatocytes were frozen in a standard freezing container. In accordance with this observation, the cultivation of cryopreserved hepatocytes from cattle was less effective than that of porcine hepatocytes. Hepatocytes from cattle can be successfully cryopreserved and partially cultured after cryopreservation but with lower percentage than porcine hepatocytes.

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Cell-based clinical and experimental methods for assisting the function of impaired livers – Present and future of liver support systems

Pluta

Ciezkowska

Wiśniewska

et al. 2021

Biocybernetics and Biomedical Engineering

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Differentiation-Promoting Medium Additives for Hepatocyte Cultivation and Cryopreservation

Cited by 5 publications

References 94 publications

Scientific Opinion of the Scientific Panel on Plant Protection Products and their Residues (PPR Panel) on testing and interpretation of comparative in vitro metabolism studies

Scientific Opinion of the Scientific Panel on Plant Protection Products and their Residues (PPR Panel) on testing and interpretation of comparative in vitro metabolism studies

Comparative cryopreservation of bovine and porcine primary hepatocytes

Cell-based clinical and experimental methods for assisting the function of impaired livers – Present and future of liver support systems

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