High-throughput quantification of the effect of DMSO on the viability of lung and breast cancer cells using an easy-to-use spectrophotometric trypan blue-based assay

Abstract

One of the main aspects investigated in potential therapeutic compounds is their effect on cells viability and proliferative ability. Although various methods have been developed to investigate these aspects, these methods present with shortcomings in terms of either cost, availability, accuracy, precision, or throughput. This study describes a simple, economic, reproducible, and high-throughput assay to quantify cell death and proliferation. In this assay, adherent cells are fixed, stained with trypan blue, and measured for trypan blue internalization using a spectrophotometric absorbance plate reader. Corresponding cell counts to the absorbance measurements are extrapolated from a standard curve. This assay was used to measure the effect of dimethyl sulfoxide (DMSO) on the viability of breast and lung cancer cells. Decrease in cell count associated with the increase in DMSO percentage and exposure time. The assay’s results closely correlated with the conventional trypan blue exclusion assay (Pearson correlation coefficient ( r ) > 0.99; p < 0.0001), but with higher precision. The assay developed in this study can be used for various applications such as optimization, cell treatment investigations, proliferation, and cytotoxicity studies.

RETRACTED: Autophagy regulates the Wnt/GSK3β/β-catenin/cyclin D1 pathway in mesenchymal stem cells (MSCs) exposed to titanium dioxide nanoparticles (TiO2NPs)

“…Cell viability in MSCs exposed to TiO 2 NPs or control was determined by after the cells were stained with trypan blue [ 35 ]. Preparation for cell counting was conducted as described in a previous study [ 36 ].…”

Anti-Proliferative and Pro-Apoptotic Effects ofCalligonum comosum(L’Her.) Methanolic Extract in Human Triple-Negative MDA-MB-231 Breast Cancer Cells

“… 30 The TNBC cells (0.25 × 10 6 /cm 2 , unless otherwise mentioned) were seeded in tissue culture plates (Corning Inc., Corning, NY, USA). The next day, the cells were exposed or not exposed to 1.8% DMSO (concentration detected in 200 μg/ml CcME and reported to be non-toxic, 31 ) or different concentrations (25-200 µg/mL) of CcME for 24 h incubation.…”

Untitled

“…Roswell Park Memorial Institute (RPMI) medium was prepared by mixing 40 mL of RPMI, 10 mL of fetal bovine serum (FBS) and 200 µL Antibiotic / antimycotic (Antibiotic antimycotic solution with Streptomycin 10 mg/20 mL, 10,000 U Penicillin, Amphotericin B and 0.9% normal saline). About 1 mL of RPMI medium was dispensed into falcon tubes, added 30 µL of Phytohemagglutinin (PHA), 100 µL of hPBMCs, and incubated with an atmosphere of 95% air and 5% CO 2 at 37 ºC for 4 h. Before incubation, counting the number of cells from 10 µL 15,16 .…”

“…Trypan Blue Dye Exclusion Method: About 10 µl of cells with 10 µl of Trypan blue (0.4% Trypan blue in a balanced salt solution) and cell suspension were mixed (1:1) and left for 10 min at room temperature. Transferred 10 µl of this onto hemocytometer, and observed under a microscope for viability count 16 . Percent viability was calculated by applying the following formula.…”