Design, alpha testing, and beta testing of a 3-D printed open-hardware portable cryopreservation device for aquatic species

Efforts in development of germplasm repositories to preserve genetic resources of aquatic species are impeded globally by a lack of standardized, inexpensive, reproducible, and portable cryopreservation technologies. The present work demonstrates a 3-D printed standardizable freezing device that can be used with nitrogen vapor shipping dewars for on-site sperm cryopreservation for aquatic species and be distributed as open-hardware. This device could hold 22 French straws (0.25-mL or 0.5-mL) and a quick-release ring design could eject straws directly into a canister inside a dewar by pressing a button after freezing. The final prototypes produced cooling rates of 1 to 68 degrees C/min for 0.25-mL straws, and 3 to 37 degrees C/min for 0.5-mL straws with a material cost of 3.5 USD for a single device and 1,820-2,562 USD for batch production of 20 replicates (including labor and purchase of 3-D printers). Progressing through design, prototyping, and testing was delineated to help guide the development of open-source devices within cryopreservation user communities.