Three-Dimensional Printing of Vitrification Loop Prototypes for Aquatic Species

Area of Science:

• Cryopreservation and Materials Science
• Biotechnology and Bioengineering

Background:

• Vitrification is a rapid cryopreservation technique forming amorphous solids, typically in small volumes.
• Current vitrification methods can be limited by device availability and standardization.

Purpose of the Study:

• To design and fabricate novel, open vitrification devices using 3D printing.
• To evaluate the impact of device geometry and loading methods on vitrification efficiency.

Main Methods:

• Utilized 3D Computer-Aided Design (CAD) to create elliptical loop designs.
• Fabricated prototype devices using acrylonitrile butadiene styrene (ABS) and polylactic acid (PLA) filaments on MakerBot® 3D printers.
• Assessed vitrification quality based on visual classification (crystalline ice vs. amorphous solid) after loading via pipetting or submersion.

Main Results:

• Pipetting offered more reliable film formation, while submersion was faster with fewer filling issues.
• Loop configurations yielding higher vitrification quality were generally longer and had fewer layers (lower height).
• Device geometry significantly influenced heat transfer and subsequent vitrification success.

Conclusions:

• 3D printing facilitates the standardization and customization of vitrification devices.
• Optimized loop dimensions and loading techniques enhance cryopreservation outcomes.
• This approach offers a flexible platform for advancing vitrification research and applications.