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
This method describes a very reliable and robust in vitro model for the screening of the cholestatic liability of drugs and other chemical entities. The 3D spheroids generated from primary human hepatocytes can be cultivated up to 28 days, allowing long-term exposures which can depict otherwise

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

Organisation: Vrije Universiteit Brussel (VUB)
Status: Still in development
The method detects two facets of drug-induced cytotoxicity i.e. the intracellular accumulation of phospholipids and of neutral lipids, i.e. phospholipidosis and steatosis respectively. The assay makes use of a kit containing an aqueous, red-fluorescent formulation of labelled phospholipids (LipidTOX

Last updated on: 28-02-2022 - 10:58

Contact: Kaat Leroy
Organisation: Vrije Universiteit Brussel (VUB)
Status: History of use
This method assesses general cytotoxicity. Upon disruption of the cell membrane, lactate dehydrogenase (LDH) is released. LDH catalyzes the interconversion of pyruvate and lactate with concomitant interconversion of reduced (NADH) and oxidized (NAD+) nicotinamide adenine dinucleotide. The principle

Last updated on: 28-02-2022 - 10:58

Contact: Kaat Leroy
Organisation: Vrije Universiteit Brussel (VUB)
Status: History of use
Human embryonic kidney (HEK) 293 FT cells is a celline that is very easy to culture and is used to obtain high viral titers. “293” is a reference to the 293th experiment wherein the cell line was discovered. A transfection with an adenovirus type 5 DNA fragment took place, causing the cell line to

Last updated on: 28-02-2022 - 10:58

Contact: Matthias Rombaut
Organisation: Vrije Universiteit Brussel (VUB)
Status: History of use
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
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
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,…)
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
The in vitro Alkaline Comet Assay is a microgel electrophoresis technique which allows to measure DNA damage (single and double strand breaks, alkali labile sites, incomplete excision repair sites and cross links) cell by cell. Cells are mixed with 0.8% Low Melting Point Agarose which is spread as a

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

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