The present standard procedure describes a protocol for measuring the urea concentration in supernatant of human stem cell-derived hepatocyte-like cells. This procedure relies on a chromogenic reagent that forms a colored complex specifically with urea. The latter can be measured and is directly
Last updated on: 28-02-2022 - 10:59
Contact: Cannot be disclosed
Organisation: Vrije Universiteit Brussel (VUB)
Status: History of use, Internally validated
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
Contact: Vânia Vilas-Boas
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
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
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
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
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 monocyte-activation test (MAT) is used to detect or quantify substances that activate human monocytes or monocytic cells to release endogenous mediators such as pro-inflammatory cytokines, for example tumour necrosis factor alpha (TNFα), interleukin-1 beta (IL-1β) and interleukin-6 (IL-6). These
Last updated on: 22-02-2022 - 10:56
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
