Despite the accelerating extinction crisis, fewer than 50 taxa (7%) of U.S. Threatened and endangered animals had cell lines cryopreserved prior to the launch of our informed biobanking project…

Despite the accelerating extinction crisis, fewer than 50 taxa (7%) of U.S. Threatened and endangered animals had cell lines cryopreserved prior to the launch of our informed biobanking project pilot in december 2022. Tissue samples have been preserved with various methods, many of which have yet to be validated, and therefore some tissues may not be viable for future applications. Cell lines are the gold standard for biobanking. There are universal methods for generating cell lines for mammals and birds from tissue biopsies collected in a non-lethal manner. Cryopreservation and thaw methods are universal, and celllines maintain the highest viability rates of any biomaterial. Our vision with the informed biobanking program was to develop a scalable biobanking workflow that ensures all U.S. Endangered species can be biobanked as part of long-term recovery strategies. We recognize this is an ambitious aspiration requiring new science and technology for the full diversity of endangered species, but for mammals and birds biobanking cell lines can be carried out now. For amphibians and reptiles, taxa for which cell lines are difficult to produce currently, tissue samples can be cryopreserved with the best present day standardized methods so that in the future, when the science has advanced, cell lines can be established. Project purpose. Our purpose is to continue generating cell lines and biobanking living tissue samples for threatened and endangered taxa for USFWS region 2, inclusive of at-risk and proposed listed taxa of major concern to the region project objectives. The desired outcome is to establish at least 1 male and 1 female cell line for as many of the mammals and birds listed in table 1 as possible given successful tissue collections and available funding. Cell lines will be stored long-term at the USDA germplasm biorepository. For amphibians and reptiles, the goal will be to biobank living tissues (AKA express tissue banking) for 1 female and 1 male, and also store those tissues long-term at the USDA germplasm biorepository. We recognize IT may not be possible to collect tissues from all the taxa listed in table 1 over the course of a single year due to the rarity and elusiveness of some taxa, as well as unforeseen circumstances that may impede collection (e.g. inclement weather, wildfire, etc.). Therefore, our goal is to establish cell lines from tissues on a first come, first serve basis until available funds are exhausted. The aim of establishing cell lines and express tissue banking for all taxa will be a longer-term goal, dependent on available funds in the following years. Methodology. Region 2 USFWS staff and their permitted academic and NGO partners working in the field and or captive propagation facilities will be responsible for collecting tissues for biobanking, following the available protocols. Revive restore will coordinate with species leads, staff, and partners to supply teams with sampling kits customized for their species needs and the necessary forms for transport and metadata recording. Samples will be shipped to viagen for cell culture generation or express tissue banking. When ready, samples will be shipped to the USDA germplasm repository for long-term storage. Revive restore will update USFWS staff throughout the process as to the status of each tissue sample. Tissue biopsies will be purposely collected from live mammals and birds during standard field monitoring activities or medical exams procedures for captive individuals. Biobanking efforts can also salvage tissues opportunistically from individuals found deceased if the carcass is found before the tissues have degraded, or from individuals taken (euthanized) for permitted recovery activities or other justified circumstances. Sampling of reptiles and amphibians will be restricted to post-mortem tissue sampling adhering to the conditions stated above.