I am trying to run a cell viability assay with LN229, but these cells are cultured in DMEM which contain phenol red. Is the MTS assay sensitive to the additional of phenol red in the media? If so, is there another colormetric assay I could do (guessing MTT will have a similar problem to MTS)?
Dear Min!
I carry out MTT analysis on glioma cells grown in DMEM medium with phenol red. Phenolic red does not affect MTT results.
Article Di-Ethylhexylphthalate (DEHP) Modulates Cell Invasion, Migra...
2.2. Cell Culture Glioblastoma cancer cell line, LN-229, was purchased from American Type Culture Collection (Rockville, MD, USA). The cells were cultured and maintained in DMEM containing 10% fetal bovine serum, 1% penicillin/streptomycin, and 2% glutamine. LN-229 cell lines were grown in BD primaria tissue culture dishes, with dimensions of 100 × 20 mm at 37 °C with 5% CO2 in a humidifier incubator and carried at 2.0 × 106 cells/mL, passaging two to three times weekly as needed.
2.6. Cell Proliferation Assay Cell proliferation was indirectly assessed with a colorimetric, (3-(4,5-dimethylthiazol-2-yl)-5- (3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) assay obtained from Promega (Madison, WI, USA). Cells (100 µL, number = 5,000) were plated on 96 well plates (Thermo Fisher Scientific, Waltham, MA, USA) and infected with shS100P and/or treated with 2.5 μg/µL DEHP. After 24 h of incubation, DMEM medium was removed and followed by the addition of 20 µL of MTS solution to each well. The 96 well plates were placed in an incubator at 37 °C in 5% CO2. The absorbance of the solution was measured at 490 nm in one hour increments for three hours using a spectrophotometer (Bio-Rad Model 550; Bio-Rad Laboratories, Inc., Hercules, CA, USA).
Yes, you can confidently perform you experiment. The presence of phenol red in DMEM will not affect your results.
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