Summary: An assessment and prediction tool of a drug-induced toxicity on human retinoblastoma cells.
Model Description
An assessment and prediction of study compound-induced toxicity on human retinoblastoma cells (WERI-Rb-1) in vitro.
| Cells | WERI-Rb-1 cells1-2 (RIKEN, Japan) |
| Viability assessment | WERI-Rb-1 cells are grown on 96-well plates. The cells can be exposed to testing molecules or formulations for selected times (e.g. 4h, 24h). Cytotoxicity is measured using resazurin reduction assay3. A cell permeable redox indicator resazurin (7-Hydroxy-3H-phenoxazin-3-one-10-oxide) is used to measure oxidation-reduction reactions in viable cells. The non-fluorescent resazurin is irreversibly reduced by metabolically active cells to highly fluorescent metabolite resorufin (7-Hydroxy-3H-phenoxazin-3-one). Resorufin formation is measured by using Cytation 3 multi-mode reader (BioTek Instruments, Winooski, VT, USA). The quantity of resorufin produced is proportional to the number of viable cells. |
| Positive control | tert-butyl hydroperoxide (tBHP) at concentrations 0-30 mM |
| Read-outs | 1. Cell viability (%) 2. IC50 value (50% cell viability) for positive control 3. IC50 value is calculated for study compound if applicable |
Outcomes and Read-Outs
References
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- McFall RC, Nagy RM, Nagle BT, McGreevy LM. Scanning electron microscopic observation of two retinoblastoma cell lines. Cancer Res. 1978,38(9):2827-2835.
- Nociari et al. A novel one-step, highly sensitive fluorometric assay to evaluate cell-mediated cytotoxicity. J Immunol Methods. 1998; 213(2):157-167. PMID: 9692848 DOI: 10.1016/s0022-1759(98)00028-3