Propidium iodide staining method for testing the cytotoxicity of 2,4,6-trichlorophenol and perfluorooctane sulfonate at low concentrations with Vero cells

Abstract

In this article, the feasibility of propidium iodide (PI) staining based on cell membrane damage as a sensitive and quantitative method to test the cytotoxicity of typical lipophilic compounds including 2,4,6-trichlorophenol (TCP) and perfluorooctane sulfonate (PFOS) was examined. The sensitivity of PI staining was compared to that of the methylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT) assay. We found a good correlation between PI uptake and increasing concentrations of TCP (10–50 μM) and PFOS (20–50 μM) at lower concentrations than those between growth inhibition ratio determined by the MTT assay and increasing concentrations of TCP (150–400 μM) and PFOS (100–500 μM). These findings indicated that the PI staining was more sensitive than the MTT assay. Furthermore, both the amount of PI uptake and the ratio of morphologically changed cells increased with increasing TCP concentrations, suggesting that PI staining may be a suitable quantitative substitute for the original method based on the observation of morphological changes in Vero cells. Our data also suggest that it is possible that the cell membrane damage that resulted in increased PI uptake may be due to variations in the phospholipid and protein content of the membrane, which are affected by interactions between the lipophilic compounds and components of the cell membrane.

Keywords:

• Vero cells
• cell membrane damage
• PI staining
• lipophilic compounds

Acknowledgments

The authors thank China Natural Science Foundation for supporting to this work (No. 20677043 and 40871217).