Drug-Induced Hepatotoxicity Assay Utiilizing upcyte Hepatocytes

The principal application of this assay is to rapidly screen test compounds to identify those that induce acute liver cell toxicity. This kit includes two aliquots of cryopreserved Luminescent Reporter Hepatocytes (upcyte(R)), donor 10-13, isolated from an adult Caucasian female, that have been further modified to constitutively express the luciferase enzyme. The level of luciferase activity expressed in the cells is dependent on the complex coordination of normal cellular processes, including the coupled rates of energy metabolism, transcription, translation, and the turnover of their respective mRNA and protein macromolecules. The hepatocytes are prepared using INDIGO's proprietary CryoMite(TM) process, which yields high cell viability post-thaw, and provides the convenience of immediately dispensing cells into assay plates. There is no need for intermediate treatment steps such as spin-and-rinse of cells, viability determinations, or cell titer adjustments prior to assay setup. An overnight culture period allows full recovery of the post-thaw hepatocytes and the establishment of a confluent cell monolayer that is ready to receive the user's test compounds. In addition to two aliquots of the reporter hepatocytes, the kit provides two cell culture-ready assay plates, optimized Cell Culture Media (CCM) for use in all steps of the assay procedure (cell thawing, seeding, and preparation of the treatment media), luciferase detection reagent, and a reference compound that provides a positive control for hepatotoxicity. The reagents and materials provided in this assay kit are formatted to allow the user to choose between two alternative assay setups. In one scenario 48 culture wells may be setup at two different times. In the other assay scenario 96 culture wells may be setup at one time.