Untangling the Gordian Knot of Aplysia sea hare egg masses: An integrated open-hardware system for standardized egg strand sizing and packaging for cryopreservation research and application

Abstract

The California sea hare (Aplysia californica) provides a powerful biomedical model system for studying aspects of neurological development and damage, behavior, aging, and hypoxia. Aplysia encapsulate their zygotes within strands that result in tangled egg masses that greatly complicate culture and experimentation. The historical and current importance of Aplysia for biomedical research and the mounting climate crisis necessitates protection of Aplysia genetic resources. The goal of this work was to prototype open-hardware sizing, processing, and packaging devices for A. californica early life stages suitable for integration into a cryopreservation pathway. The Strand Centi-Sizer was a low-cost, fused filament fabrication 3-D printable device that increased experiment preparation efficiency and standardized the cutting of egg strands customizable to user needs. A downstream system of 3-D printed devices was also prototyped to address inefficiencies in handling of egg strand sections for processing and packaging into existing cryopreservation straw platforms. Time studies were conducted comparing manual methods (i.e., no specialized equipment) with open hardware to demonstrate utility of the devices and to encourage community members to design and prototype new devices to address recurrent and novel problems in other aquatic animals that produce egg strands. Improvements in design could further increase efficiency, standardization, and reproducibility, and extend the application of these devices to other research communities, such as shrimp or salamander spermatophores, sea anemone body part (e.g., pedal lacerate) cryopreservation, or study areas such as vitrification.