Modified method for evaluation of plasma membrane integrity in eukaryotic cell

Area of Science:

• Microbiology
• Cell Biology
• Biotechnology

Background:

• Accurate enumeration of viable cells is crucial in various biological and clinical applications.
• Existing methods for cell viability assessment can be complex, time-consuming, or require specialized equipment.
• There is a need for a simple, cost-effective method to evaluate cell viability.

Purpose of the Study:

• To develop and evaluate a novel method for assessing the number of viable cells based on dye absorption by dead cells.
• To investigate the efficacy of myramistin as a pore-forming agent in this viability assay.
• To assess the impact of myramistin on the viability of Candida albicans, Malassezia sympodialis, and human epitheliocytes.

Main Methods:

• A method was developed utilizing bromcresol purple dye absorption by dead cells from the incubation medium.
• Myramistin was employed as a pore-forming agent to facilitate dye entry into dead cells.
• The correlation between live cell numbers and the percentage of dye absorbed was analyzed.

Main Results:

• A clear inverse correlation was observed between the number of live cells and the percentage of bromcresol purple dye absorbed.
• Myramistin treatment (100 µg/ml for 2 hours) significantly reduced viable Candida albicans and Malassezia sympodialis cells by 2 and 1 order of magnitude, respectively.
• Human epitheliocytes absorbed a comparable amount of dye to Candida cells under identical myramistin treatment conditions.

Conclusions:

• The proposed method offers a simple and equipment-free approach for evaluating viable cell numbers.
• Bromcresol purple dye absorption by dead cells, facilitated by myramistin, serves as a reliable indicator of cell viability.
• Myramistin demonstrates potent cell-killing activity against relevant fungal pathogens and affects human epithelial cells similarly.