The 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 gel

Last updated on: 20-03-2019 - 17:30

Contact: Roel Anthonissen
Organisation: Sciensano
Status: History of use

The in vitro micronucleus test is a genotoxicity test for the detection of micronuclei in the cytoplasm of interphase cells. Micronuclei may originate from acentric chromosome fragments (i.e. lacking a centromere), or whole chromosomes that are unable to migrate to the poles during the anaphase

Last updated on: 20-03-2019 - 17:26

Organisation: Sciensano
Status: Published in peer reviewed journal, Validated by an external party (e.g. OECD, EURL ECVAM,…)

The RT-qPCR assay is used to identify genotoxic and non-genotoxic compounds. Herefore metabolic-competent human HepaRG cells are exposed to the IC10 value (measured by the MTT test). A microassay is performed to select 84 genes that show the most robust rates of correct classification of the

Last updated on: 20-03-2019 - 17:22

Organisation: Sciensano
Partners: Vrije Universiteit Brussel
Status: Internally validated, Published in peer reviewed journal

The Ames test is a short-term bacterial reverse mutation assay specifically designed to detect a wide range of chemical substances that can produce genetic damage that leads to permanent gene mutations. The test employs several histidine dependent Salmonella strains each carrying different

Last updated on: 20-03-2019 - 17:16

Contact: Roel Anthonissen
Organisation: Sciensano
Status: Published in peer reviewed journal, 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. It accumulates in the lysosomes. Dying

Last updated on: 20-03-2019 - 17:11

Contact: Roel Anthonissen
Organisation: Sciensano
Status: History of use

Cultivated liver cells are fixed with 4 % (w/v) paraformaldehyde (PFA) for 10 minutes at room temperature and subsequently incubated for 15 minutes with 100 mM glycin solution, used to saturate reactive groups generated after PFA fixation. These cells are subsequently incubated for 10 minutes

Last updated on: 20-03-2019 - 16:22

Contact: Joery De Kock
Organisation: Vrije Universiteit Brussel
Status: History of use, Internally validated, Published in peer reviewed journal

Rat liver epithelial cells are cultivated at 100 % confluency on 100 µg/mL rat tail collagen type I coated culture dishes in base medium and sequentially exposed to hepatogenic growth factors and cytokines. Base medium consisted of William’s E medium without glutamine supplemented with 7.33 IE/mL

Last updated on: 20-03-2019 - 16:18

Contact: Joery De Kock
Organisation: Vrije Universiteit Brussel
Status: History of use, Internally validated, Published in peer reviewed journal

Using this method you can measure the (relative) lipid load in human skin-derived stem cells differentiated towards hepatic cells. This method could also be applied on other cell types (e.g. HepG2), since it is based on the following publication: "M. T. Donato et al., Chem. Biol. Interact. 181,

Last updated on: 20-03-2019 - 16:07

Contact: Joost Boeckmans
Organisation: Vrije Universiteit Brussel
Status: Internally validated

Mononuclear cells (MNC) are isolated from bone marrow aspirates by density gradient centrifugation and washed in Hank's buffered salt solution. MNC are seeded at a cell density of 2 × 10E4 cells/cm2 in low glucose DMEM supplemented with 15% (v/v) heat-inactivated FBS, 2 mM L-glutamine and 0.5% (v

Last updated on: 20-03-2019 - 16:01

Contact: Joery De Kock
Organisation: Vrije Universiteit Brussel
Status: History of use, Internally validated, Published in peer reviewed journal

Approximately 125 g of processed adipose tissue is incubated for 90 minutes at 37 °C in dissociation medium (1:1) consisting of 1% (v/v) bovine serum albumin and 1 mg/mL collagenase A in phosphate buffered saline (PBS). After two filtration steps, the filtrate is carefully brought on top of 15 mL

Last updated on: 20-03-2019 - 15:57

Contact: Joery De Kock
Organisation: Vrije Universiteit Brussel
Status: History of use, Internally validated, Published in peer reviewed journal