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: 08-11-2019 - 09:32

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

Human skin-derived precursor cells (hSKP) between are cultivated at 90% confluency on 100 µg/mL collagen type I in base medium and sequentially exposed to hepatogenic factors. Base medium consists of Dulbecco’s Modified Eagle Medium + GLUTAMAX / F12 Nutrient supplement (3:1), supplemented with 7

Last updated on: 08-11-2019 - 09:31

Contact: Cannot be disclosed
Organisation: Vrije Universiteit Brussel
Status: History of use, Internally validated, Published in peer reviewed journal

E. coli is one of the organisms of choice for the production of recombinant proteins. DH5 alpha cells are commonly used for maintenance, propagation and mutation, whilst BL21(DE3) and C43(DE3) are mainly used for expression of the transgene. The advantage of C43(DE3) is that is used to produce

Last updated on: 08-11-2019 - 09:28

Organisation: Vrije Universiteit Brussel
Status: Still in development, History of use, Internally validated

Freshly collected human foreskin samples are incubated with 0.2 mg/mL Liberase DH solution and incubated overnight at 4°C. The next day, the epidermis is removed and the tissue is incubated at 37°C for another 10-20 minutes, depending on the sample size. After processing the samples, typically 5

Last updated on: 08-11-2019 - 09:27

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

HeLa cells are the first continuous cancer cell line and were isolated from the aggressive glandular cervical cancer of a 31-year old woman. It was the first aneuploid line derived from human tissue maintained in continuous cell culture. Knowledge of almost every process that occurs in human

Last updated on: 08-11-2019 - 09:25

Organisation: Vrije Universiteit Brussel
Status: Still in development, History of use

This method describes the steps to go from a liver to a decellularized matrix. It uses mild and strong detergents to destroy cells and keep the extracellular matrix intact. This matrix can then be used for a variety of purposes, including (but not limited to) repopulation, basis for coating and

Last updated on: 08-11-2019 - 09:24

Contact: Paul Claes
Organisation: Vrije Universiteit Brussel
Status: Published in peer reviewed journal

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: 08-11-2019 - 09:22

Contact: Paul Claes
Organisation: Vrije Universiteit Brussel

Human skin-derived adult stem cells differentiated towards hepatic cells (hSKP-HPC) are used in this method (R. M. Rodrigues et al., Stem Cells Dev. 23, 44–55 (2014)). These cells are exposed to a cocktail of insulin and glucose at certain concentrations. After 24h of exposure, these cells

Last updated on: 08-11-2019 - 09:19

Contact: Joost Boeckmans
Organisation: Vrije Universiteit Brussel
Status: Still in development

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

Last updated on: 08-11-2019 - 09:18

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

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: 08-11-2019 - 09:18

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