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Cell Cytotoxicity Assay

MP Manitosh Pandey
AS Alok Kumar Singh
RT Ritesh Thakare
ST Sakshi Talwar
PK Pratiksha Karaulia
AD Arunava Dasgupta
SC Sidharth Chopra
AP Amit Kumar Pandey
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Cell toxicity was performed against Vero cell using the MTT assay. ~103 cells/well were seeded in 96 well plate and incubated at 37 °C with an 5% CO2 atmosphere. After 24 h, compound was added ranging from 100–12.5 mg/L and incubated for 72 h at 37 °C with an 5% CO2 atmosphere. After the incubation was over, MTT was added at 5 mg/L in each well, incubated at 37 °C for further 4 hours, residual medium was discarded, 0.1 mL of DMSO was added to solubilise the formazan crystals and OD was taken at 540 nm for the calculation of CC50. CC50 is defined as the lowest concentration of compound which leads to a 50% reduction in cell viability. Doxorubicin was used as positive control and each experiment was repeated in triplicate.

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