{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,26]],"date-time":"2025-11-26T21:28:52Z","timestamp":1764192532908,"version":"3.41.2"},"reference-count":85,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2021,5,27]],"date-time":"2021-05-27T00:00:00Z","timestamp":1622073600000},"content-version":"vor","delay-in-days":26,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Current Protocols"],"published-print":{"date-parts":[[2021,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Protocols for cryopreservation of mouse embryos and sperm are important for preserving genetically engineered mice (GEMs) used in research to study human development and diseases. Embryo cryopreservation is mainly carried out using either of two protocols: controlled gradual cooling or vitrification. Sperm cryopreservation protocols include two methodologies that are commonly referred to as JAX and CARD. Quality\u2010control measures are necessary to ensure that GEMs are properly cryopreserved so that they can be retrieved for future use. An archiving system is also important in keeping proper records of frozen sperm and embryos. Frozen embryos and sperm are now preferred over live mice for shipping to distant locations. This article describes detailed protocols used in cryopreservation of mouse embryos and sperm, as well as their retrieval to live mice. \u00a9 2021 U.S. Government.<\/jats:p><jats:p>This article was corrected on 28 July 2022. See the end of the full text for details.<\/jats:p><jats:p><jats:bold><jats:italic>Sperm cryopreservation<\/jats:italic><\/jats:bold><\/jats:p><jats:p><jats:bold>Basic Protocol 1<\/jats:bold>: JAX protocol for sperm cryopreservation<\/jats:p><jats:p><jats:bold>Support Protocol 1<\/jats:bold>: JAX protocol for making sperm cryopreservation medium<\/jats:p><jats:p><jats:bold>Basic Protocol 2<\/jats:bold>: JAX protocol for IVF of mouse sperm<\/jats:p><jats:p><jats:bold>Alternate Protocol 1<\/jats:bold>: Modified CARD protocol for sperm cryopreservation<\/jats:p><jats:p><jats:bold>Support Protocol 2: CARD protocol for<\/jats:bold> making sperm cryopreservation medium<\/jats:p><jats:p><jats:bold>Alternate Protocol 2<\/jats:bold>: CARD protocol for IVF of mouse sperm<\/jats:p><jats:p><jats:bold><jats:italic>Embryo cryopreservation<\/jats:italic><\/jats:bold><\/jats:p><jats:p><jats:bold>Basic Protocol 3<\/jats:bold>: Cryopreserving and thawing 2\u2010cell embryos<\/jats:p><jats:p><jats:bold>Alternate Protocol 3<\/jats:bold>: Cryopreserving and thawing 8\u2010cell to morula\u2010stage embryos<\/jats:p><jats:p><jats:bold><jats:italic>Surgical transfer of embryos<\/jats:italic><\/jats:bold><\/jats:p><jats:p><jats:bold>Basic Protocol 4<\/jats:bold>: Infundibulum transfer of 2\u2010cell to morula\u2010stage embryos<\/jats:p>","DOI":"10.1002\/cpz1.138","type":"journal-article","created":{"date-parts":[[2021,5,27]],"date-time":"2021-05-27T16:25:03Z","timestamp":1622132703000},"source":"Crossref","is-referenced-by-count":3,"title":["Cryopreservation Protocols for Genetically Engineered Mice"],"prefix":"10.1002","volume":"1","author":[{"given":"Glenn","family":"Longenecker","sequence":"first","affiliation":[{"name":"National Institute of Dental and Craniofacial Research National Institutes of Health  Bethesda Maryland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kyoungin","family":"Cho","sequence":"additional","affiliation":[{"name":"National Institute of Allergy and Infectious Diseases National Institutes of Health  Bethesda Maryland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jaspal S.","family":"Khillan","sequence":"additional","affiliation":[{"name":"National Institute of Allergy and Infectious Diseases National Institutes of Health  Bethesda Maryland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ashok B.","family":"Kulkarni","sequence":"additional","affiliation":[{"name":"National Institute of Dental and Craniofacial Research National Institutes of Health  Bethesda Maryland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2021,5,27]]},"reference":[{"key":"e_1_2_17_2_1_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-6828-2_1"},{"key":"e_1_2_17_2_2_1","doi-asserted-by":"publisher","DOI":"10.1071\/bi9510581"},{"key":"e_1_2_17_2_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/170326aO"},{"key":"e_1_2_17_2_4_1","first-page":"8","article-title":"Frozen sperm as an alternative to shipping live mice","volume":"42","author":"Awasthi P. 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