DES In Cell Cultures: Assessing the Cytotoxicity of An Alternative 3D Cell Culture Model

Primary Author

Isiah Arambula

Faculty Mentor

Dr. Kelly Nash

Additional Mentor(s)

Alejandro Morales Betancourt


Deep Eutectic Solvents are a relatively new class of solvent with green properties that place them in a positive light. However, these solvents still require further experimentation and assessment to determine the precise applications they could be best used for. One of those potential applications is the cell culture. DES could be used to create a 3D culture as an alternative to other 3D models. These models are inexpensive compared to murine models and would still bypass the limitations of 2D. There is conflicting information among literature mentioning both toxic and proliferation effects of cultured cells. The focus of this project is to determine the cytotoxicity of relevant DES, that we plan to expand usage on within our lab. To test the cytotoxicity, ARPE-19 cells were cultured until being seeded into 96 well plates at a desired concentration per well. After 24-hour incubation, the cells were treated with different dilutions of DES. Among these solvents, are Choline Chloride(ChCl)-Urea, ChCl-Sucrose-Water, and ChCl-Glycerol. The cells were placed back to incubate for another 24 hours before adding a reagent containing Luciferase to measure Luminescence in the microplate reader. Data was collected and exported to Excel for analysis. Results showed no significant increase in viability against the measured controls. As expected, the different solvents have different points in concentration where lethality increases rapidly. The DES showed extreme cytotoxicity at our highest concentration and decreased at lower concentrations. Future directions will be to explore other concentrations and molar ratios of DES. With enough data and conclusions, we want to expand to 3D cancer cell cultures and utilize nanoparticle-based treatments on these cancer cells.


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