1. For each test time point, remove medium from the top of each gel.
  2. Add 1ml fresh medium onto the sample and next add 100 µl of AlamarBlue solution.
  3. Place the gels in the incubator at 37°C for 2 hours.
  4. Pipette and transfer 200 µl of the supernatant to 96-well plate.
  5. To ensure the accuracy of reading, the average value from triplicates should be calculated.
  6. To ensure accuracy of readings, the average value from triplicates should be calculated using a Fluorescence plate reader FLUOstar OPTIMA (BMG, Labtech, UK) with excitation wavelength at 530 nm and emission wavelength at 590 nm.
  7. Statistical analyses performed using the One-way ANOVA test.