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Measuring the Densities of Aqueous Glasses at Cryogenic Temperatures
Published on: June 28, 2017
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High-throughput evaluation of cryoprotective agents for mixture effects that reduce toxicity
Nima Ahmadkhani1, Cameron Sugden1, James D Benson2
1School of Chemical, Biological and Environmental Engineering, Oregon State University, US.
Cryobiology
|September 17, 2025
Summary
Discovering less toxic cryoprotective agents (CPAs) is key for organ vitrification. This study screened 21 compounds, finding four binary CPA mixtures significantly reduced toxicity, enhancing cell viability for better cryopreservation.
Area of Science:
- Biotechnology
- Cell Biology
- Cryobiology
Background:
- Vitrification enables cryopreservation of organs but requires high concentrations of toxic cryoprotective agents (CPAs).
- Limited CPA options hinder the development of safer vitrification solutions.
- Novel CPAs are needed to improve efficacy and reduce toxicity.
Purpose of the Study:
- To screen 21 compounds for toxicity individually and in binary combinations.
- To identify novel cryoprotective agent (CPA) mixtures with reduced toxicity.
- To develop a high-throughput method for assessing CPA toxicity.
Main Methods:
- High-throughput screening of 21 compounds at room temperature.
- Toxicity assessment using bovine pulmonary artery endothelial cells (BPAEC).
- Evaluation of individual compounds and binary combinations at various concentrations and exposure durations.
Main Results:
- Toxicity increased with CPA concentration and exposure time.
- Four binary CPA combinations (formamide/glycerol, DMSO/1,3-propanediol, 1,2-propanediol/diethylene glycol, 1,3-propanediol/diethylene glycol) showed significantly reduced toxicity.
- The 6 mol/kg mixtures of these binary combinations resulted in higher cell viability compared to single CPA solutions.
Conclusions:
- Binary CPA mixtures can significantly reduce toxicity compared to individual CPAs.
- A high-throughput screening method aids in building a CPA toxicity database.
- This approach supports the development of predictive models for novel, low-toxicity CPA mixtures for vitrification.
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