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 exhibit a
Last updated on: 08-11-2019 - 09:19
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: 08-11-2019 - 09:17
The neutral red uptake assay is a cell viability assay that allows in vitro quantification of xenobiotic-induced cytotoxicity. The assay relies on the ability of living cells to incorporate and bind neutral red, a weak cationic dye, in lysosomes. As such, cytotoxicity is expressed as a concentration
Last updated on: 08-11-2019 - 09:17
Contact: Robim Rodrigues
Organisation: Vrije Universiteit Brussel
Status: Published in peer reviewed journal, Validated by an external party (e.g. OECD, EURL ECVAM,…)
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: 08-11-2019 - 09:17
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: 08-11-2019 - 09:15
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: 08-11-2019 - 09:15
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: 08-11-2019 - 09:14
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: 08-11-2019 - 09:14
Contact: Cannot be disclosed
Organisation: Vrije Universiteit Brussel
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: 08-11-2019 - 09:14
Contact: Cannot be disclosed
Organisation: Vrije Universiteit Brussel
Status: History of use, Internally validated
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: 08-11-2019 - 09:14
