Isolation of rat hepatocytes

This method describes the steps from a living rat to a single cell solution of primary hepatocytes. This requires surgery on the lab animal, a perfusion with buffer solution, a digestion with collagenase and a filtration step to obtain primary hepatocytes.

Last updated on: 28-02-2022 - 11:01

Contact: Paul Claes
Organisation: Vrije Universiteit Brussel (VUB)
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Measurement of the extracellular release of adenosine triphosphate in cultured primary rat hepatocytes

ATP transports chemical energy within cells by serving as a substrate for kinases and as such fulfills a vital function in numerous cellular processes such as cell injury and subsequent cell death. ATP is therefore a crucial player in these events that are results of intracellular stress.

Last updated on: 28-02-2022 - 10:59

Contact: Axelle Cooreman
Organisation: Vrije Universiteit Brussel (VUB)
Status: History of use
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Cytotoxicity measurement in cultured primary rat hepatocytes

The MTT test is performed to determine the in vitro cytotoxicity of selected chemicals. The mitochondrial enzyme succinate deydrogenase is responsible for the biotransformation of toxic agents and MTT. The ability of cells to reduce MTT provides an indication of the mitochondrial integrity and

Last updated on: 28-02-2022 - 10:59

Contact: Axelle Cooreman
Organisation: Vrije Universiteit Brussel (VUB)
Status: History of use
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The detection of cholestasis-inducing agents in cultured primary rat hepatocytes

The standard operating procedure describes a method to assess the cholestasis-inducing potential of chemicals, in casu in cultures of primary rat hepatocytes. The procedure relies on the accumulation of the fluorescent bile salt export pump (Bsep) substrate cholyl-lysyl-fluorescein (CLF) in the

Last updated on: 28-02-2022 - 10:59

Contact: Raf Van Campenhout
Organisation: Vrije Universiteit Brussel (VUB)
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Sudan Red III in situ staining of cultured primary rat hepatocytes

The standard operating procedure for Sudan Red III in situ staining of cultured rat hepatocytes describes how to detect one of the aspects of drug-induced cytotoxicity i.e. the intracellular accumulation of lipids or in other words steatosis, in primary rat hepatocyte cultures. It is based on the

Last updated on: 28-02-2022 - 10:59

Contact: Raf Van Campenhout
Organisation: Vrije Universiteit Brussel (VUB)
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Measurement of reactive oxygen species in cultured primary rat hepatocytes

Basically, the standard operating procedure outlined in this document consists of the following steps, namely, (i) preparation of the cells for the DCFH-DA assay, (ii) DCFH-DA test procedure, (iii) processing of the results. Practical details are provided for each of these steps and are followed by

Last updated on: 28-02-2022 - 10:59

Contact: Cannot be disclosed
Organisation: Vrije Universiteit Brussel (VUB)
Status: History of use, Internally validated
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Multiplex immunoassay for quantification of antigens of diphtheria, tetanus and acellular pertussis in human combined DTaP vaccines

The multiplex immunoassay is based on the Luminex technology and allows the detection of diphtheria, tetanus and acellular pertussis antigens of human combined vaccines in the same run. As potency test of these vaccines are currently performed on animal through challenge and/or serological assays,

Last updated on: 28-02-2022 - 09:19

Contact: Maxime Vermeulen
Organisation: Sciensano
Partners: VAC2VAC
Status: Still in development
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Innovative two-chamber skin explant model to study skin diseases in marine fish

Maaike Vercauteren developed the innovative in vitro 'two-chamber skin explant model'. Pieces of skin are kept and examined in a controlled laboratory environment. This is no sinecure, because the skin must continue to function as if it were still attached to the fish. However, the tested setup

Last updated on: 22-02-2022 - 15:13

Contact: Cannot be disclosed
Organisation: Instituut voor Landbouw-, Visserij- en Voedingsonderzoek (ILVO), Ghent University (UGent)
Status: Internally validated
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Direct Peptide Reactivity Assay

The DPRA is an in chemico method which quantifies the remaining concentration of cysteine- or lysine-containing peptide following 24 hours incubation with the test chemical at 25 +/-2,5ºC. The synthetic peptides contain phenylalanine to aid in the detection. Relative peptide concentration is

Last updated on: 22-02-2022 - 10:59

Contact: Bart Desmedt
Organisation: Sciensano
Status: History of use, Internally validated, Validated by an external party (e.g. OECD, EURL ECVAM,…)
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Neutral Red Uptake Assay

The neutral red uptake (NRU) assay provides a quantitative measurement of the number of viable cells. The test is based on the ability of living cells to take up and bind neutral red (NR), a dye which easily penetrates cell membranes via non-ionic diffusion and accumulates in the lysosomes. Dying

Last updated on: 21-02-2022 - 15:23

Contact: Roel Anthonissen
Organisation: Sciensano
Status: History of use, Published in peer reviewed journal
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