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University Provost\u2019s Fund","award":["LAB94420"],"award-info":[{"award-number":["LAB94420"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Animals"],"abstract":"<jats:p>The axolotl (Ambystoma mexicanum) draws great attention around the world for its importance as a biomedical research model, but housing and maintaining live animals is increasingly expensive and risky as new transgenic lines are developed. The goal of this work was to develop an initial practical pathway for sperm cryopreservation to support germplasm repository development. The present study assembled a pathway through the investigation of axolotl sperm collection by stripping, refrigerated storage in various osmotic pressures, cryopreservation in various cryoprotectants, and in vitro fertilization using thawed sperm. By the stripping of males, 25\u2013800 \u00b5L of sperm fluid was collected at concentrations of 1.6 \u00d7 106 to 8.9 \u00d7 107 sperm\/mL. Sperm remained motile for 5 d in Hanks\u2019 Balanced Salt Solution (HBSS) at osmolalities of 100\u2013600 mOsm\/kg. Sperm cryopreserved in 0.25 mL French straws at 20 \u00b0C\/min in a final concentration of 5% DMFA plus 200 mM trehalose and thawed at 25 \u00b0C for 15 s resulted in 52 \u00b1 12% total post-thaw motility. In six in vitro fertilization trials, 20% of eggs tested with thawed sperm continued to develop to stage 7\u20138 after 24 h, and a third of those embryos (58) hatched. This work is the first report of successful production of axolotl offspring with cryopreserved sperm, providing a general framework for pathway development to establish Ambystoma germplasm repositories for future research and applications.<\/jats:p>","DOI":"10.3390\/ani14020206","type":"journal-article","created":{"date-parts":[[2024,1,9]],"date-time":"2024-01-09T03:36:58Z","timestamp":1704771418000},"page":"206","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Establishment of a Practical Sperm Cryopreservation Pathway for the Axolotl (Ambystoma mexicanum): A Community-Level Approach to Germplasm Repository Development"],"prefix":"10.3390","volume":"14","author":[{"given":"Nicholas","family":"Coxe","sequence":"first","affiliation":[{"name":"Aquatic Germplasm and Genetic Resources Center, School of Renewable Natural Resources, Louisiana State University Agricultural Center, Baton Rouge, LA 70820, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"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":[{"vocabulary":"crossref","role":"author"}]},{"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":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1324-6873","authenticated-orcid":false,"given":"Rose","family":"Upton","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":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9057-2771","authenticated-orcid":false,"given":"Sarah","family":"Bodenstein","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"},{"name":"Louisiana Sea Grant College Program, Louisiana State University, Baton Rouge, LA 70803, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven Randal","family":"Voss","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, Ambystoma Genetic Stock Center and Spinal Cord and Brain Injury Research Center, University of Kentucky, Lexington, KY 40536, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria T.","family":"Gutierrez-Wing","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":[{"vocabulary":"crossref","role":"author"}]},{"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":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,1,8]]},"reference":[{"key":"ref_1","unstructured":"(2023, October 19). 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