Assessment of toxicity responses in human retinoblastoma cells 

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 the 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 

Fig. 1. A, B) Typical morphology of WERI-Rb-1 cells showing rosettes (R) and chain (C) formation.
Fig. 1. A, B) Typical morphology of WERI-Rb-1 cells showing rosettes (R) and chain (C) formation. Scale bar 100 µm. C, D) Characteristic grape-like clusters of WERI-Rb-1 cells at higher cell density. Scale bar 100 µm (left) and 200 µm (right). 

References

  • McFall RC, Sery TW, Makadon M. Characterization of a new continuous cell line derived from a human retinoblastoma. Cancer Res. 1977,37(4):1003-1010.
  • 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

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