Title

Ethanol as a solvent in pharmaceutical toxicity testing should be used with caution

Document Type

Poster

Campus where you would like to present

Ellensburg

Event Website

https://digitalcommons.cwu.edu/source

Start Date

18-5-2020

Abstract

Mouse liver cells have a vital role in many scientific fields as they are used to test for toxicity of pharmaceutical compounds. Additionally, ethanol is usually used as the vehicle control to dissolve those compounds. Different concentrations of ethanol were investigated to find the optimal amount to be used for cells grown in vitro. Vector transfected Hepa-V mouse liver cells were grown in 25 cm2 flasks using a complete media solution containing DMEM/F12, Nu-Serum and 1% Penicillin-Streptomycin. After reaching 80% confluency, the cells were exposed to either the vehicle control (0.83 to 3.34% of ethanol) or the xenobiotics (5 to 20 μM of Antimycin A) for 24 hours. WST-8 was added to the cells and absorbance of the solution was measured at 450 nm using Synergy 2 plate reader. The readouts showed a decreasing trend in the amount of living cells in both final solutions of vehicle control treatment and xenobiotics treatment. However, those treated with just ethanol observed a more linear relationship (R2 = 0.96) compared to those exposed to Antimycin A alone (R2 = 0.68). Cell viability of cells treated with Antimycin A reached a threshold after 10 μM while those introduced to ethanol experienced more cell death as percent of ethanol in the vehicle control increased. This results confirm that ethanol has a noticeable effect on cell viability and it should be used no higher than 2.5% in cell culture.

Faculty Mentor(s)

Carin Thomas

Department/Program

Chemistry

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May 18th, 12:00 PM

Ethanol as a solvent in pharmaceutical toxicity testing should be used with caution

Ellensburg

Mouse liver cells have a vital role in many scientific fields as they are used to test for toxicity of pharmaceutical compounds. Additionally, ethanol is usually used as the vehicle control to dissolve those compounds. Different concentrations of ethanol were investigated to find the optimal amount to be used for cells grown in vitro. Vector transfected Hepa-V mouse liver cells were grown in 25 cm2 flasks using a complete media solution containing DMEM/F12, Nu-Serum and 1% Penicillin-Streptomycin. After reaching 80% confluency, the cells were exposed to either the vehicle control (0.83 to 3.34% of ethanol) or the xenobiotics (5 to 20 μM of Antimycin A) for 24 hours. WST-8 was added to the cells and absorbance of the solution was measured at 450 nm using Synergy 2 plate reader. The readouts showed a decreasing trend in the amount of living cells in both final solutions of vehicle control treatment and xenobiotics treatment. However, those treated with just ethanol observed a more linear relationship (R2 = 0.96) compared to those exposed to Antimycin A alone (R2 = 0.68). Cell viability of cells treated with Antimycin A reached a threshold after 10 μM while those introduced to ethanol experienced more cell death as percent of ethanol in the vehicle control increased. This results confirm that ethanol has a noticeable effect on cell viability and it should be used no higher than 2.5% in cell culture.

https://digitalcommons.cwu.edu/source/2020/COTS/36