ASSESSMENT OF ON- AND OFF-TARGET EFFECTS USING AN IPS CARDIOMYOCYTE CELL MODEL Evaluating cell health is an important step in the process of drug discovery screening. Test compounds must be evaluated for the desired effects on the viability of target cells and for undesirable toxic off-target effects. Induced Pluripotent Stem (iPS) cells are an emerging tool in a variety of areas, including toxicity screening. Cardiotoxicity is a lethal side effect of drug treatment that has resulted in the removal of therapies from the market for safety reasons. To demonstrate the applicability of iPS-derived cardiomyocytes as an off-target model for toxicity screening, we conducted parallel experiments using a target human K562 cancer cell line model alongside the iPS cardiomyocyte cell line. Select drugs known to target cancerous cells were tested, and the toxicity profiles between the two cell types compared. To assess overall cell health and cytotoxicity, we used the ApoTox-Glo™ Triplex Assay, a combination of chemistries used to measure viability, cytotoxicity and apoptosis. The results presented here show how the ApoTox-Glo™ Assay can be used with different cell types to assess various compounds for on-target and off-target effects . 14 The tyrosine kinase inhibitor Imatinib, targets the mutant bcr-abl kinase found ApoTox-Glo is a trademark of Promega Corporation. Cardiomyocyte K562 The tyrosine kinase inhibitor Imatinib shows off-target cytotoxic effects in iPS cardiomyocytes . in leukocytes of patients with Chronic Myelogenous Leukemia (CML). The immortalized K562 cell line was isolated from a patient with CML, and serves as a model system for the development of cancer therapies targeting this disease. Here, a serial titration of Imatinib was applied to each cell line and incubated for 24 hours at 37°C, 5% CO2. Following treatment, the ApoTox-Glo™ Assay was performed. APPLICATION NOTE
Scoor meer met een web winkel in uw magazines. Velen gingen u voor en publiceerden jaarverslagen online.
Promega Benelux eNews - February 2012 Lees publicatie 10Home