Feb 24, 2025
Resazurin cell viability assay in hiPSC-derived ineurons
- 1UCL / The Francis Crick Institute
Protocol Citation: Laura Mahoney 2025. Resazurin cell viability assay in hiPSC-derived ineurons. protocols.io https://dx.doi.org/10.17504/protocols.io.5jyl8d4r6g2w/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: February 24, 2025
Last Modified: February 24, 2025
Protocol Integer ID: 123276
Abstract
This protocols describes the steps to follow in order to assess cell viability in hiPSC-derived ineurons cultured in 96-well plates via resazurin.
Resazurin preparation and addition to cells
Resazurin preparation and addition to cells
A stock solution was prepared at a concentration of 10mg/ml by dissolving resazurin dye (7-hydroxy-3H-phenoxazin-3-one 10-oxide) in dH2O.
Day 14 ineurons were cultured in 200 µL culture media without antioxidants. Ferroptosis inducer treatments were performed for 24 hours.
To asses viability, 20 µL of the resazurin stock solution was directly added to the each well of a 96-well plate containing 200 µL of culture media (10% of cell culture volume per well – final concentration 100mg/ml) and plate was return to the 37 °C incubator for 02:00:00
Resazurin readout
Resazurin readout
Following the 2h resazurin incubation, samples were analyzed fluorometrically on a microplate reader (The SPARK‱ Multimode microplate reader, Ex = 540nM, Em = 600nM). Background signals obtained from cell-free wells were subtracted from each sample.
Cell viability under treatment conditions were reported as a percentage relative to untreated control cells.