{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T06:00:14Z","timestamp":1781848814216,"version":"3.54.5"},"reference-count":23,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2025,5,14]],"date-time":"2025-05-14T00:00:00Z","timestamp":1747180800000},"content-version":"vor","delay-in-days":13,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"publisher","award":["306749\/2021"],"award-info":[{"award-number":["306749\/2021"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Reprod Domestic Animals"],"published-print":{"date-parts":[[2025,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>High concentrations of cryoprotectants required for testicular vitrification result in a toxic environment for cells. To mitigate this issue, a suitable alternative is to combine cryoprotectants. The temperature for warming a vitrified sample is also important to assure cell viability. Thus, the aim of this work was to evaluate how combining cryoprotectants (ethylene glycol\u2010EG, glycerol\u2010GLY, and dimethyl sulfoxide\u2010DMSO) in pairs and using two warming temperatures (37\u00b0C and 50\u00b0C) influence cellular morphology, tubular morphometry, and mitochondrial activity after testicular vitrification of dogs. Testicular fragments from ten adult dogs were distributed among the fresh control group (CTR) and the experimental groups according to the combination of cryoprotectants and temperatures (EG\/GLY37, EG\/GLY50, DMSO\/GLY37, DMSO\/GLY50, DMSO\/EG37 and DMSO\/EG50). The fragments were vitrified in a final concentration of 5.6\u2009mol\/L (2.8\u2009mol\/L of each of the cryoprotectants combined two by two) and subsequently warmed up to 37\u00b0C\/30\u2009s or 50\u00b0C\/5\u2009s. Following this, they were processed for histomorphological, morphometric, and mitochondrial activity evaluations with Rhodamine 123. In the morphometric evaluation, all vitrified groups showed a significant reduction in tubular diameter (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.05). All experimental groups showed greater basement membrane separation when related to the CTR (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.05). DMSO\/EG37 showed the greatest basement membrane separation when compared to all other groups (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.05). Regarding membrane retraction, all vitrified groups, regardless of the warming temperature, had greater retraction when related to CTR (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.05), except DMSO\/GLY50, which did not differ from any group (<jats:italic>p<\/jats:italic>\u2009&gt;\u20090.05). Regarding the distinction between spermatogonia and Sertoli cells, no groups warmed up to 50\u00b0C differed from the control, except DMSO\/GLY37. For nuclear visualisation, none of the vitrified groups differed from the CTR (<jats:italic>p<\/jats:italic>\u2009&gt;\u20090.05), except DMSO\/GLY37 (<jats:italic>p<\/jats:italic>\u2009&lt;\u20090.05), which showed better nuclear visualisation. For the nuclear condensation parameter, there were no significant differences among the groups (<jats:italic>p<\/jats:italic>\u2009&gt;\u20090.05). Mitochondrial activity was reduced in all vitrified samples, regardless of the combination of cryoprotectants and warming temperature (<jats:italic>p<\/jats:italic>\u2009&gt;\u20090.05). It was concluded that the association of DMSO\/GLY50 presented better preservation of morphological aspects.<\/jats:p>","DOI":"10.1111\/rda.70074","type":"journal-article","created":{"date-parts":[[2025,5,14]],"date-time":"2025-05-14T09:25:35Z","timestamp":1747214735000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Influence of Three Combinations of Cryoprotectants and Two Warming Temperatures on Cellular Morphology, Morphometry and Mitochondrial Activity of Vitrified Canine Testicles"],"prefix":"10.1111","volume":"60","author":[{"given":"Juliana","family":"de Souza Fernandes","sequence":"first","affiliation":[{"name":"Laborat\u00f3rio de Reprodu\u00e7\u00e3o de Carn\u00edvoros (LRC) Universidade Estadual do Cear\u00e1 (UECE)  Fortaleza CE Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00e9ssyka Ara\u00fajo","family":"Noronha","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Reprodu\u00e7\u00e3o de Carn\u00edvoros (LRC) Universidade Estadual do Cear\u00e1 (UECE)  Fortaleza CE Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francisco Denilson Rodrigues","family":"Gomes","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Manipula\u00e7\u00e3o de O\u00f3citos e Fol\u00edculos Ovarianos Pr\u00e9\u2010Antrais (LAMOFOPA) Universidade Estadual do Cear\u00e1 (UECE)  Fortaleza CE Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bruna Farias","family":"Brito","sequence":"additional","affiliation":[{"name":"Centro Universit\u00e1rio Fametro (UNIFAMETRO)  Fortaleza CE Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gisele Karla Sena","family":"Guimar\u00e3es","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Reprodu\u00e7\u00e3o de Carn\u00edvoros (LRC) Universidade Estadual do Cear\u00e1 (UECE)  Fortaleza CE Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Herlon Victor Rodrigues","family":"Silva","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Reprodu\u00e7\u00e3o de Carn\u00edvoros (LRC) Universidade Estadual do Cear\u00e1 (UECE)  Fortaleza CE Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Leda Maria Costa Pereira","family":"Bersano","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Reprodu\u00e7\u00e3o de Carn\u00edvoros (LRC) Universidade Estadual do Cear\u00e1 (UECE)  Fortaleza CE Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9793-1968","authenticated-orcid":false,"given":"L\u00facia Daniel Machado","family":"da Silva","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Reprodu\u00e7\u00e3o de Carn\u00edvoros (LRC) Universidade Estadual do Cear\u00e1 (UECE)  Fortaleza CE Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2025,5,14]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cryobiol.2021.01.010"},{"issue":"4","key":"e_1_2_10_3_1","first-page":"32","article-title":"Avan\u00e7os Tecnol\u00f3gicos da Criopreserva\u00e7\u00e3o de Ov\u00f3citos","volume":"1","author":"Barbosa R. 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