The genetic resources of aquatic biomedical organisms are the products of millions of years of evolution, decades of scientific development, and hundreds of millions of dollars of research funding investment. Aquatic organisms have become powerful models in biomedical research and laboratories around the world have produced tens of thousands of mutant and transgenic lines. Maintaining these valuable genotypes as live animals is expensive, risky, and beyond the capacity of most stock centers. As such, cryopreservation has become a necessity and typically these genetic resources are now maintained as samples of inconsistent quality frozen with rudimentary techniques. Quality control has not been practiced in any systematic way, reproducibility is poor, and protocols are not standardized. It is common to have problems and failures in fertilization resulting in lost lines that need to be recreated, causing facilities to waste considerable time and effort. This is largely due to the false notion that neglecting quality control saves time and money. However, rather than being reduced, these costs are shifted downstream through wasted storage space and reduced fertilization. This pervasive lack of reliability and reproducibility has placed the substantial investments in biomedical research at great risk. These pervasive problems were the subject of a 2017 NIH Cryopreservation Workshop to develop germplasm repositories to protect aquatic biomedical genetic resources. This proposal directly responds to the specific needs identified by research communities and the Directors of the five NIH-funded aquatic animal stock centers at that workshop. The mechanism identified to deliver the much-needed research and capacity development was through establishment of a Hub and repository network based on the model of the AGGRC, which is specifically directed at translating research into applied practice.
The Specific Aims of this proposal are to: 1) Establish comprehensive repository systems at NIH-funded stock centers for sperm of Xenopus frogs and Ambystoma salamanders. 2) Establish a comprehensive repository system at the NIH-funded stock center for early life stages (e.g., embryos, larvae) of Aplysia sea hares and required algae species. 3) Establish a comprehensive, centralized unit (?Hub?) to integrate activities across NIH-funded stock centers and their communities, and develop approaches and documentation for cryopreservation. This includes development of protocols and pathways, outreach programs, community interaction, standardization, freezing services, and training. The AGGRC was developed to directly address these needs and is uniquely suited to perform and integrate the necessary research, stock center, and network-level activities.
Aquatic models are rapidly transforming genetic research of human disease, and as thousands of new research lines become available each year, we must develop the abilities and resources necessary to reliably preserve them in repositories to protect their valuable genetic resources. The most significant problems constraining repository development are the lack of reliability and efficiency in cryopreservation caused by the absence of quality control, reproducibility, and standardization which prevent the coupling and integration of activities among research laboratories and stock centers, jeopardize maintenance of valuable lines, and produce costly inefficiency and duplication of efforts that slow research. Given that these perils multiply as the pool of cryopreserved samples is expanded, we propose to develop appropriate technology through research for amphibians (i.e., Ambystoma and Xenopus) and invertebrates (i.e., Aplysia), and provide routine access to reproducibility, training, and standardization as community-level approaches through development of a centralized repository resource Hub and establishment of a repository network to address all aquatic animal biomedical models.
