{"id":9840,"date":"2025-08-16T16:41:58","date_gmt":"2025-08-16T16:41:58","guid":{"rendered":"http:\/\/drmhasghari.com\/interspecies-somatic-cell-nuclear-transfer-in-asiatic-cheetah-using-nuclei-derived-from-post-mortem-frozen-tissue-in-absence-of-cryo-protectant-and-in-vitro-matured-domestic-cat-oocytes\/"},"modified":"2025-09-12T11:42:34","modified_gmt":"2025-09-12T11:42:34","slug":"interspecies-somatic-cell-nuclear-transfer-in-asiatic-cheetah-using-nuclei-derived-from-post-mortem-frozen-tissue-in-absence-of-cryo-protectant-and-in-vitro-matured-domestic-cat-oocytes","status":"publish","type":"post","link":"https:\/\/drmhasghari.com\/en\/interspecies-somatic-cell-nuclear-transfer-in-asiatic-cheetah-using-nuclei-derived-from-post-mortem-frozen-tissue-in-absence-of-cryo-protectant-and-in-vitro-matured-domestic-cat-oocytes\/","title":{"rendered":"Interspecies somatic cell nuclear transfer in Asiatic cheetah using nuclei derived from post-mortem frozen tissue in absence of cryo-protectant and in vitro matured domestic cat oocytes"},"content":{"rendered":"<div class=\"wpb-content-wrapper\"><p>[vc_row][vc_column][vc_empty_space][vc_column_text css=&#8221;&#8221;]<\/p>\n<div id=\"ez-toc-container\" class=\"ez-toc-v2_0_85 ez-toc-wrap-center counter-hierarchy ez-toc-counter-rtl ez-toc-custom ez-toc-container-direction\">\n<div class=\"ez-toc-title-container\">\n<p class=\"ez-toc-title\" style=\"cursor:inherit\">Table of Contents<\/p>\n<span class=\"ez-toc-title-toggle\"><a href=\"#\" class=\"ez-toc-pull-right ez-toc-btn ez-toc-btn-xs ez-toc-btn-default ez-toc-toggle\" aria-label=\"Toggle Table of Content\"><span class=\"ez-toc-js-icon-con\"><span class=\"\"><span class=\"eztoc-hide\" style=\"display:none;\">Toggle<\/span><span class=\"ez-toc-icon-toggle-span\"><svg style=\"fill: #62c0e0;color:#62c0e0\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\" class=\"list-377408\" width=\"20px\" height=\"20px\" viewBox=\"0 0 24 24\" fill=\"none\"><path d=\"M6 6H4v2h2V6zm14 0H8v2h12V6zM4 11h2v2H4v-2zm16 0H8v2h12v-2zM4 16h2v2H4v-2zm16 0H8v2h12v-2z\" fill=\"currentColor\"><\/path><\/svg><svg style=\"fill: #62c0e0;color:#62c0e0\" class=\"arrow-unsorted-368013\" xmlns=\"http:\/\/www.w3.org\/2000\/svg\" width=\"10px\" height=\"10px\" viewBox=\"0 0 24 24\" version=\"1.2\" baseProfile=\"tiny\"><path d=\"M18.2 9.3l-6.2-6.3-6.2 6.3c-.2.2-.3.4-.3.7s.1.5.3.7c.2.2.4.3.7.3h11c.3 0 .5-.1.7-.3.2-.2.3-.5.3-.7s-.1-.5-.3-.7zM5.8 14.7l6.2 6.3 6.2-6.3c.2-.2.3-.5.3-.7s-.1-.5-.3-.7c-.2-.2-.4-.3-.7-.3h-11c-.3 0-.5.1-.7.3-.2.2-.3.5-.3.7s.1.5.3.7z\"\/><\/svg><\/span><\/span><\/span><\/a><\/span><\/div>\n<nav><ul class='ez-toc-list ez-toc-list-level-1 eztoc-toggle-hide-by-default' ><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-1\" href=\"https:\/\/drmhasghari.com\/en\/interspecies-somatic-cell-nuclear-transfer-in-asiatic-cheetah-using-nuclei-derived-from-post-mortem-frozen-tissue-in-absence-of-cryo-protectant-and-in-vitro-matured-domestic-cat-oocytes\/#Highlights\" >Highlights<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-2\" href=\"https:\/\/drmhasghari.com\/en\/interspecies-somatic-cell-nuclear-transfer-in-asiatic-cheetah-using-nuclei-derived-from-post-mortem-frozen-tissue-in-absence-of-cryo-protectant-and-in-vitro-matured-domestic-cat-oocytes\/#Abstract\" >Abstract<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-3\" href=\"https:\/\/drmhasghari.com\/en\/interspecies-somatic-cell-nuclear-transfer-in-asiatic-cheetah-using-nuclei-derived-from-post-mortem-frozen-tissue-in-absence-of-cryo-protectant-and-in-vitro-matured-domestic-cat-oocytes\/#Introduction\" >Introduction<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-4\" href=\"https:\/\/drmhasghari.com\/en\/interspecies-somatic-cell-nuclear-transfer-in-asiatic-cheetah-using-nuclei-derived-from-post-mortem-frozen-tissue-in-absence-of-cryo-protectant-and-in-vitro-matured-domestic-cat-oocytes\/#Discussion\" >Discussion<\/a><\/li><li class='ez-toc-page-1 ez-toc-heading-level-2'><a class=\"ez-toc-link ez-toc-heading-5\" href=\"https:\/\/drmhasghari.com\/en\/interspecies-somatic-cell-nuclear-transfer-in-asiatic-cheetah-using-nuclei-derived-from-post-mortem-frozen-tissue-in-absence-of-cryo-protectant-and-in-vitro-matured-domestic-cat-oocytes\/#Acknowledgments\" >Acknowledgments<\/a><\/li><\/ul><\/nav><\/div>\n<h2 data-start=\"473\" data-end=\"487\"><span class=\"ez-toc-section\" id=\"Highlights\"><\/span>Highlights<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<ul data-start=\"488\" data-end=\"912\">\n<li data-start=\"488\" data-end=\"619\">\n<p data-start=\"490\" data-end=\"619\">Non-viable nuclei obtained from an Asiatic cheetah carcass preserved at \u2013\u06f2\u06f0\u00b0C were used for iSCNT.<\/p>\n<\/li>\n<li data-start=\"620\" data-end=\"711\">\n<p data-start=\"622\" data-end=\"711\">Cloned cheetah embryos developed in vitro up to the <strong data-start=\"673\" data-end=\"683\">morula<\/strong> stage but then arrested.<\/p>\n<\/li>\n<li data-start=\"712\" data-end=\"815\">\n<p data-start=\"714\" data-end=\"815\">Evidence of <strong data-start=\"746\" data-end=\"767\">embryonic fragmentation<\/strong> and <strong data-start=\"770\" data-end=\"802\">abnormal chromosome segregation<\/strong> was observed in cheetah iSCNT embryos.<\/p>\n<\/li>\n<li data-start=\"816\" data-end=\"912\">\n<p data-start=\"818\" data-end=\"912\"><strong data-start=\"818\" data-end=\"837\">Genomic instability<\/strong> was identified as the main cause of failure to reach the blastocyst stage.<\/p>\n<\/li>\n<\/ul>\n<h2 data-start=\"919\" data-end=\"927\"><span class=\"ez-toc-section\" id=\"Abstract\"><\/span>Abstract<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p data-start=\"928\" data-end=\"1365\">Recent advances in <strong data-start=\"954\" data-end=\"989\">somatic cell nuclear transfer<\/strong> (SCNT) have raised great hopes for preserving the genetic resources of endangered animals through ex situ conservation technologies. Most SCNT studies use live donor cells for nuclear transfer into recipient oocytes. However, in certain situations where only frozen tissues of rare animals\u2014without proper cryopreservation\u2014are available, obtaining viable cells is difficult or impossible.  <\/p>\n<p data-start=\"1367\" data-end=\"1656\">In this study, the feasibility of <strong data-start=\"1394\" data-end=\"1442\">interspecies somatic cell nuclear transfer (iSCNT)<\/strong> in the Asiatic cheetah (Acinonyx jubatus venaticus) was investigated. Nuclei were isolated from frozen tissue preserved at \u2013\u06f2\u06f0\u00b0C without cryoprotectant, and in vitro\u2013matured domestic cat oocytes were used as recipients. <\/p>\n<p data-start=\"1658\" data-end=\"1936\">No cell growth was observed from initial cultures of skin and tendon fragments or after enzymatic digestion and single-cell culture. Viability staining revealed that nearly all cells had ruptured membranes and were non-viable. Therefore, nuclei were directly injected into enucleated domestic cat oocytes.  <\/p>\n<p data-start=\"1938\" data-end=\"2214\">Early signs of nuclear reprogramming were observed \u06f2 hours after iSCNT and significantly increased by \u06f4 hours. The percentages of iSCNT reconstructions that developed to the \u06f4\u2013\u06f1\u06f6 cell and morula stages were \u06f3\u06f2.\u06f3 \u00b1 \u06f7.\u06f3%, \u06f1\u06f8.\u06f2 \u00b1 \u06f9.\u06f8%, and \u06f5.\u06f9 \u00b1 \u06f4.\u06f3%, respectively. However, none of the embryos progressed to the blastocyst stage.  <\/p>\n<p data-start=\"2216\" data-end=\"2409\">In the control group using cat cells and cat oocytes, cleavage rates (\u06f5\u06f1.\u06f3 \u00b1 \u06f1\u06f3.\u06f9% and \u06f7\u06f7.\u06f3 \u00b1 \u06f4.\u06f0%) and blastocyst formation (\u06f1\u06f1.\u06f3 \u00b1 \u06f3.\u06f3% and \u06f1\u06f6.\u06f8 \u00b1 \u06f3.\u06f8%) were comparable to standard results.<\/p>\n<p data-start=\"2411\" data-end=\"2661\">Overall, this study demonstrated that domestic cat oocytes could partially reprogram non-viable nuclei from the Asiatic cheetah and return them to early embryonic stages. This is the first report of iSCNT in the Asiatic cheetah using frozen non-viable cells. <\/p>\n<h2 data-start=\"2668\" data-end=\"2676\"><span class=\"ez-toc-section\" id=\"Introduction\"><\/span>Introduction<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p data-start=\"2677\" data-end=\"2868\">Earth\u2019s current biodiversity is the product of over \u06f3.\u06f5 billion years of evolution. However, in the past century, human activities have caused rapid biodiversity loss, placing many species at the brink of extinction. <\/p>\n<p data-start=\"2870\" data-end=\"3164\">While in situ conservation plays a vital role in species survival, new technologies such as <strong data-start=\"2968\" data-end=\"3003\">SCNT<\/strong> allow scientists to utilize genetic resources to reproduce rare species. To date, SCNT has resulted in successful cloning in more than \u06f2\u06f0 species. <\/p>\n<p data-start=\"3166\" data-end=\"3407\">Cloning an animal species requires appropriate donor cells, recipient oocytes, and surrogate mothers. For extinct or critically endangered species, this is nearly impossible, as live donor cells are unavailable and genomic integrity in frozen tissues is often compromised. <\/p>\n<p data-start=\"3409\" data-end=\"3764\">Nevertheless, recent studies have shown that even tissues frozen at very low temperatures (\u2013\u06f2\u06f0\u00b0C to \u2013\u06f8\u06f0\u00b0C) without cryoprotectant can preserve parts of genomic integrity for years. Moreover, iSCNT has previously been successfully used to produce embryos or offspring in animals such as the <strong data-start=\"3674\" data-end=\"3735\">wild cat, gray wolf, gaur, mouflon, and Pyrenean ibex. <\/strong> <\/p>\n<p data-start=\"3766\" data-end=\"4052\">The Asiatic cheetah (Acinonyx jubatus venaticus), one of the rarest subspecies and the only surviving cheetah in Iran, is currently listed as <strong data-start=\"3918\" data-end=\"3940\">critically endangered<\/strong> by the IUCN Red List. Its population is estimated to be fewer than \u06f1\u06f1\u06f0 individuals, with road accidents accounting for about \u06f4\u06f0% of deaths. <\/p>\n<p data-start=\"4054\" data-end=\"4239\">In this study, tissues were obtained from a cheetah that died in a road accident in Yazd Province on May \u06f2\u06f7, \u06f2\u06f0\u06f1\u06f0, and whose carcass had been stored at \u2013\u06f2\u06f0\u00b0C for \u06f1\u06f0 days, in order to assess the feasibility of cloning.<\/p>\n<h2 data-start=\"4246\" data-end=\"4252\"><span class=\"ez-toc-section\" id=\"Discussion\"><\/span>Discussion<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p data-start=\"4253\" data-end=\"4555\">This study represents the first attempt to generate interspecies cloned embryos of the Asiatic cheetah in vitro using nuclei extracted from frozen tissue without cryoprotectant. The results showed that although cell division up to the early stages was possible, genomic instability prevented further development to the blastocyst stage. <\/p>\n<p data-start=\"4557\" data-end=\"4758\">These findings mark an important step toward understanding the capabilities and limitations of iSCNT in conserving endangered species and provide prospects for future use of this method in preserving the genetic resources of rare animals. <\/p>\n<h2 data-start=\"4765\" data-end=\"4775\"><span class=\"ez-toc-section\" id=\"Acknowledgments\"><\/span>Acknowledgments<span class=\"ez-toc-section-end\"><\/span><\/h2>\n<p data-start=\"4776\" data-end=\"4983\">The authors dedicate this study to the memory of the late <strong data-start=\"4807\" data-end=\"4855\">Hosseini Sh., former head of the Yazd Province Department of Environment <\/strong>(who passed away on December \u06f3\u06f1, \u06f2\u06f0\u06f1\u06f3), and to all Iranian rangers who sacrifice their lives to protect nature and wildlife.<\/p>\n<blockquote>\n<p data-start=\"1678\" data-end=\"1785\"><strong>Link: <a href=\"https:\/\/www.sciencedirect.com\/science\/article\/abs\/pii\/S0093691X16305647\" target=\"_blank\" rel=\"noopener\">Interspecies somatic cell nuclear transfer in Asiatic cheetah using nuclei derived from post-mortem frozen tissue in absence of cryo-protectant and in vitro matured domestic cat oocytes<\/a><\/strong><\/p>\n<\/blockquote>\n<p>[\/vc_column_text][vc_empty_space][\/vc_column][\/vc_row]<\/p>\n<\/div>","protected":false},"excerpt":{"rendered":"<p>Cloning of the Asiatic cheetah using interspecies somatic cell nuclear transfer (iSCNT) with frozen tissue lacking cryoprotectant and domestic cat oocytes was investigated. The results showed that the embryos developed up to the morula stage but did not reach the blastocyst stage due to genomic instability. This study represents the first scientific report of iSCNT in the Asiatic cheetah and offers a new perspective for the conservation of endangered species.  <\/p>\n","protected":false},"author":2,"featured_media":9629,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"site-sidebar-layout":"default","site-content-layout":"","ast-site-content-layout":"default","site-content-style":"default","site-sidebar-style":"default","ast-global-header-display":"","ast-banner-title-visibility":"","ast-main-header-display":"","ast-hfb-above-header-display":"","ast-hfb-below-header-display":"","ast-hfb-mobile-header-display":"","site-post-title":"","ast-breadcrumbs-content":"","ast-featured-img":"","footer-sml-layout":"","ast-disable-related-posts":"","theme-transparent-header-meta":"default","adv-header-id-meta":"","stick-header-meta":"default","header-above-stick-meta":"","header-main-stick-meta":"","header-below-stick-meta":"","astra-migrate-meta-layouts":"default","ast-page-background-enabled":"default","ast-page-background-meta":{"desktop":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"ast-content-background-meta":{"desktop":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"tablet":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""},"mobile":{"background-color":"var(--ast-global-color-5)","background-image":"","background-repeat":"repeat","background-position":"center center","background-size":"auto","background-attachment":"scroll","background-type":"","background-media":"","overlay-type":"","overlay-color":"","overlay-opacity":"","overlay-gradient":""}},"footnotes":""},"categories":[82],"tags":[],"class_list":["post-9840","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-scientific-articles"],"_links":{"self":[{"href":"https:\/\/drmhasghari.com\/en\/wp-json\/wp\/v2\/posts\/9840","targetHints":{"allow":["GET"]}}],"collection":[{"href":"https:\/\/drmhasghari.com\/en\/wp-json\/wp\/v2\/posts"}],"about":[{"href":"https:\/\/drmhasghari.com\/en\/wp-json\/wp\/v2\/types\/post"}],"author":[{"embeddable":true,"href":"https:\/\/drmhasghari.com\/en\/wp-json\/wp\/v2\/users\/2"}],"replies":[{"embeddable":true,"href":"https:\/\/drmhasghari.com\/en\/wp-json\/wp\/v2\/comments?post=9840"}],"version-history":[{"count":2,"href":"https:\/\/drmhasghari.com\/en\/wp-json\/wp\/v2\/posts\/9840\/revisions"}],"predecessor-version":[{"id":9842,"href":"https:\/\/drmhasghari.com\/en\/wp-json\/wp\/v2\/posts\/9840\/revisions\/9842"}],"wp:featuredmedia":[{"embeddable":true,"href":"https:\/\/drmhasghari.com\/en\/wp-json\/wp\/v2\/media\/9629"}],"wp:attachment":[{"href":"https:\/\/drmhasghari.com\/en\/wp-json\/wp\/v2\/media?parent=9840"}],"wp:term":[{"taxonomy":"category","embeddable":true,"href":"https:\/\/drmhasghari.com\/en\/wp-json\/wp\/v2\/categories?post=9840"},{"taxonomy":"post_tag","embeddable":true,"href":"https:\/\/drmhasghari.com\/en\/wp-json\/wp\/v2\/tags?post=9840"}],"curies":[{"name":"wp","href":"https:\/\/api.w.org\/{rel}","templated":true}]}}