I am trying to see the effect of drugs on my cellular target. Before that, I have to make sure that they are not toxic to the cells. As such, I am performing cell viability assay using alamarblue assay. This is my first time doing it.
Has someone worked with MEF´s before in a cell viability assay and could please recommend me the incubation time of the dye with MEF´s before the readout?
I read that it is different for different cell lines.
Thanks, Saif
You will need to grow and optimize your time that you are interested, using for example:
Article Generation and Culture of Mouse Embryonic Fibroblasts
Dear Mohd!
You look also at the protocol:
Article Toxic effects of resazurin on cell cultures
mouse embryonic fibroblast stem cells,
All cells were seeded at a density of 10,000 cells/ml in each well of four 96-well plates (Greiner Bio-one, Monroe, NC, USA). Each well was filled with 200 μL of medium, specific to each cell type. The plates were incubated for 3 days in a 37 °C and 5 % CO2 incubator. After the initial seeding time, the wells were changed with new media which were supplemented with concentrations of resazurin ranging from 0 to 20 % (v/v) of stock alamarBlue® solution to medium. Two of the seeded plates were then incubated at 37 °C and 5 % CO2 for 3 h, while one plate was incubated for 4 days, and the last plate was incubated for 8 days. Medium was changed every 2 days.
Article Dying endothelial cells stimulate proliferation of malignant...
Cell proliferation assay
AlamarBlue assays were performed to evaluate the proliferation of glioma cells according to the manufacturer’s instructions (Invitrogen, Carlsbad, CA, USA). Glioma cells were seeded on 24-transwell receiver plates at a density of 1.0×103 cells/well. For the coculture system, ∼1.0×105 pretreated HUVECs were seeded on each companion insert. The fluorescence intensity of the glioma cells was measured with a microplate reader (FLUOstar OPTIMA, BMG Labtech, Offenburg, Germany) after an 8-h incubation with 10% alamarBlue reagent added into each well.
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