Polycaprolactone (PCL) fiber materials are fabricated using an electrospinning method. Molecular weight of PCL is 45 000 Da (Sigma-Aldrich). The electrospinning process is performed using 18wt% PCL solution dissolved in chloroform:ethanol at a ratio of 9:1. Two high-voltage sources are used to
Last updated on: 28-02-2022 - 11:00
For the generation of the human skin-derived precursors (hSKP) spheroids, ultra-low attachment 96-well plates, cell-repellent (Greiner) are used. hSKP cultured on tissue culture plates are detached with trypple (5 minutes) and collected in a tube. After centrifugation, hSKP are counted at the Burker
Last updated on: 28-02-2022 - 10:59
Contact: Alessandra Natale
Organisation: Vrije Universiteit Brussel (VUB)
Status: Still in development
ATP transports chemical energy within cells by serving as a substrate for kinases and as such fulfills a vital function in numerous cellular processes such as cell injury and subsequent cell death. ATP is therefore a crucial player in these events that are results of intracellular stress.
Last updated on: 28-02-2022 - 10:59
The MTT test is performed to determine the in vitro cytotoxicity of selected chemicals. The mitochondrial enzyme succinate deydrogenase is responsible for the biotransformation of toxic agents and MTT. The ability of cells to reduce MTT provides an indication of the mitochondrial integrity and
Last updated on: 28-02-2022 - 10:59
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: 28-02-2022 - 10:59
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: 28-02-2022 - 10:59
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
