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T32AI074490"],"award-info":[{"award-number":["ALH: T32AI074490"]}]},{"name":"National Institutes of Health","award":["R24-OD010441"],"award-info":[{"award-number":["R24-OD010441"]}]},{"name":"National Institutes of Health","award":["R24-OD028443"],"award-info":[{"award-number":["R24-OD028443"]}]},{"name":"National Institutes of Health","award":["LAB94231"],"award-info":[{"award-number":["LAB94231"]}]},{"name":"National Institutes of Health","award":["LAB94420"],"award-info":[{"award-number":["LAB94420"]}]},{"name":"National Institutes of Health","award":["MLN: 1923259"],"award-info":[{"award-number":["MLN: 1923259"]}]},{"name":"National Institutes of Health","award":["ALH: T32AI074490"],"award-info":[{"award-number":["ALH: T32AI074490"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>Hydractinia symbiolongicarpus is an emerging model organism in which cutting-edge genomic tools and resources are being developed for use in a growing number of research fields. One limitation of this model system is the lack of long-term storage for genetic resources. The goal of this study was to establish a generalizable cryopreservation approach for Hydractinia that would support future repository development for other cnidarian species. Specific objectives were to: (1) characterize basic parameters related to sperm quality; (2) develop a generalizable approach for sperm collection; (3) assess the feasibility of in vitro fertilization (IVF) with sperm after refrigerated storage; (4) assess the feasibility of IVF with sperm cryopreserved with various sperm concentrations; (5) evaluate feasibility of cryopreservation with various freezing conditions, and (6) explore the feasibility of cryopreservation by use of a 3-D printed open-hardware (CryoKit) device. Animal husbandry and sperm collection were facilitated by use of 3-D printed open hardware. Hydractinia sperm at a concentration of 2 \u00d7 107 cells\/mL stored at 4 \u00b0C for 6 d were able to achieve 50% fertilization rate. It appeared that relatively higher sperm concentration (&gt;5 \u00d7 107 cells\/mL) for cryopreservation could promote fertilization. A fertilization rate of 41\u201369% was observed using sperm equilibrated with 5, 10, or 15% (v\/v) cryoprotectant (dimethyl sulfoxide or methanol) for 20 min, cooled at a rate of 5, 10, or 20 \u00b0C\/min from 4 \u00b0C to \u221280 \u00b0C, at a cell concentration of 108\/mL, in 0.25 mL French straws. Samples cryopreserved with the CryoKit produced a fertilization rate of 72\u201382%. Establishing repository capabilities for the Hydractinia research community will be essential for future development, maintenance, protection, and distribution of genetic resources. More broadly, these generalizable approaches can be used as a model to develop germplasm repositories for other cnidarian species.<\/jats:p>","DOI":"10.3390\/ani12192537","type":"journal-article","created":{"date-parts":[[2022,9,22]],"date-time":"2022-09-22T21:10:05Z","timestamp":1663881005000},"page":"2537","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Cryopreservation of Hydractinia symbiolongicarpus Sperm to Support Community-Based Repository Development for Preservation of Genetic Resources"],"prefix":"10.3390","volume":"12","author":[{"given":"Aidan L.","family":"Huene","sequence":"first","affiliation":[{"name":"Department of Surgery, Thomas E. Starzl Transplantation Institute, University of Pittsburgh, Pittsburgh, PA 15213, USA"},{"name":"Pittsburgh Center for Evolutionary Biology and Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA"},{"name":"Aquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA 70820, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5733-4640","authenticated-orcid":false,"given":"Jack C.","family":"Koch","sequence":"additional","affiliation":[{"name":"Aquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA 70820, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4355-0155","authenticated-orcid":false,"given":"Luc\u00eda","family":"Arregui","sequence":"additional","affiliation":[{"name":"Aquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA 70820, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3838-8277","authenticated-orcid":false,"given":"Yue","family":"Liu","sequence":"additional","affiliation":[{"name":"Aquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA 70820, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthew L.","family":"Nicotra","sequence":"additional","affiliation":[{"name":"Department of Surgery, Thomas E. Starzl Transplantation Institute, University of Pittsburgh, Pittsburgh, PA 15213, USA"},{"name":"Pittsburgh Center for Evolutionary Biology and Medicine, University of Pittsburgh, Pittsburgh, PA 15213, USA"},{"name":"Department of Immunology, University of Pittsburgh, Pittsburgh, PA 15213, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1826-2848","authenticated-orcid":false,"given":"Virginia M.","family":"Weis","sequence":"additional","affiliation":[{"name":"Department of Integrative Biology, Oregon State University, Corvallis, OR 97331, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7738-1978","authenticated-orcid":false,"given":"Terrence R.","family":"Tiersch","sequence":"additional","affiliation":[{"name":"Aquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA 70820, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,9,22]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1387\/ijdb.123502gp","article-title":"Hydractinia, a pioneering model for stem cell biology and reprogramming somatic cells to pluripotency","volume":"56","author":"Plickert","year":"2012","journal-title":"Int. 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