{"id":72,"date":"2010-06-22T18:55:00","date_gmt":"2010-06-22T18:55:00","guid":{"rendered":"https:\/\/wordpress-dev.med.unc.edu\/mmrrc\/resources\/sperm-thawing-protocol\/"},"modified":"2010-06-22T18:55:00","modified_gmt":"2010-06-22T18:55:00","slug":"sperm-thawing-protocol","status":"publish","type":"page","link":"https:\/\/www.med.unc.edu\/mmrrc\/resources\/sperm-thawing-protocol\/","title":{"rendered":"Sperm Thawing Protocol"},"content":{"rendered":"<p><!-- portlet_header --><\/p>\n<p><!-- portlet_above --><\/p>\n<p><!-- description --><\/p>\n<p class=\"lead\">\n<h3><b>Samples must be stored in liquid nitrogen (or liquid nitrogen vapor) until ready to thaw<\/b><\/h3>\n<h4>Sperm Thawing<\/h4>\n<ol>\n<li>Using forceps, grasp straw and hold in air for ~10 seconds then gradually lower into 37* water bath (10ul sperm fraction first, slowly lower until buffer fraction is in water) until ice disappears (~10 seconds).<\/li>\n<li>Remove from water and gently wipe straw dry (straw may be fractured, but sample is OK)<\/li>\n<li>Holding firmly, carefully cut off both sealed ends, beveling end with the 10ul sperm aliquot.<\/li>\n<li>Transfer only the thawed sperm suspension directly into a 500ul IVF drop (do not let larger buffer column contact media).<\/li>\n<li>Allow sperm to capacitate 40-60 minutes before adding oocytes.<\/li>\n<li>Continue with IVF as usual.<\/li>\n<\/ol>\n<h4>Reference<\/h4>\n<p>Conserving, Distributing and Managing Genetically Modified Mouse Lines by Sperm Cryopreservation G. Charles Ostermeier, Michael V. Wiles, Jane S. Farley, Robert A. Taft<\/p>\n","protected":false},"excerpt":{"rendered":"<p>Samples must be stored in liquid nitrogen (or liquid nitrogen vapor) until ready to thaw Sperm Thawing Using forceps, grasp straw and hold in air for ~10 seconds then gradually lower into 37* water bath (10ul sperm fraction first, slowly lower until buffer fraction is in water) until ice disappears (~10 seconds). Remove from water &hellip; <a href=\"https:\/\/www.med.unc.edu\/mmrrc\/resources\/sperm-thawing-protocol\/\" aria-label=\"Read more about Sperm Thawing Protocol\">Read more<\/a><\/p>\n","protected":false},"author":1369,"featured_media":0,"parent":25,"menu_order":3,"comment_status":"closed","ping_status":"closed","template":"","meta":{"_acf_changed":false,"footnotes":"","_links_to":"","_links_to_target":""},"class_list":["post-72","page","type-page","status-publish","hentry","odd"],"acf":[],"_links_to":[],"_links_to_target":[],"_links":{"self":[{"href":"https:\/\/www.med.unc.edu\/mmrrc\/wp-json\/wp\/v2\/pages\/72","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/www.med.unc.edu\/mmrrc\/wp-json\/wp\/v2\/pages"}],"about":[{"href":"https:\/\/www.med.unc.edu\/mmrrc\/wp-json\/wp\/v2\/types\/page"}],"author":[{"embeddable":true,"href":"https:\/\/www.med.unc.edu\/mmrrc\/wp-json\/wp\/v2\/users\/1369"}],"replies":[{"embeddable":true,"href":"https:\/\/www.med.unc.edu\/mmrrc\/wp-json\/wp\/v2\/comments?post=72"}],"version-history":[{"count":0,"href":"https:\/\/www.med.unc.edu\/mmrrc\/wp-json\/wp\/v2\/pages\/72\/revisions"}],"up":[{"embeddable":true,"href":"https:\/\/www.med.unc.edu\/mmrrc\/wp-json\/wp\/v2\/pages\/25"}],"wp:attachment":[{"href":"https:\/\/www.med.unc.edu\/mmrrc\/wp-json\/wp\/v2\/media?parent=72"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}