{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T05:08:33Z","timestamp":1792818513687,"version":"4.5.0"},"reference-count":69,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2023,7,6]],"date-time":"2023-07-06T00:00:00Z","timestamp":1688601600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Vet. Sci."],"abstract":"<jats:p>In this study, it was hypothesized that the addition of an appropriate concentration of Y-27632 (a ROCK inhibitor) to the freezing extender prevents cryopreservation-induced apoptosis and improves embryonic development after <jats:italic>in vitro<\/jats:italic> fertilization (IVF). Semen samples were collected from five fertile Simmental bulls using an artificial vagina twice a week for 4\u2009weeks. Selected samples were pooled and diluted with Tris-egg-yolk-glycerol (TEYG) extender containing different concentrations of Y-27632 (0, 10, 20, 30, and 40\u2009\u03bcM) and then frozen in liquid nitrogen. After thawing, computer-assisted semen analysis (CASA), plasma membrane integrity, and acrosome intactness were evaluated in terms of morphological abnormalities, intracellular generation of reactive oxygen species (ROS), DNA fragmentation, phosphatidylserine (PS) externalization, and apoptotic-related gene expression. Finally, groups of frozen and thawed spermatozoa were used for bovine oocyte IVF. The results show that the semen extender at a concentration of 20\u2009\u03bcM Y-27632 effectively improved total motility (TM), curvilinear velocity (VCL), as well as the plasma membrane and acrosome integrity compared to the control group (<jats:italic>p<\/jats:italic>\u2009&amp;lt;\u20090.05). Intracellular ROS levels were significantly (<jats:italic>p<\/jats:italic>\u2009&amp;lt;\u20090.05) lower in samples treated with 30\u2009\u03bcM Y-27632 compared to the control specimen. Furthermore, supplementation of the semen extender with 20\u2009\u03bcM Y-27632 resulted in more viable spermatozoa compared with the control group (<jats:italic>p<\/jats:italic>\u2009&amp;lt;\u20090.05). According to qRT-PCR results, the expression levels of <jats:italic>BAX<\/jats:italic> and <jats:italic>CASPASE-9<\/jats:italic> genes in samples treated with 30\u2009\u03bcM Y-27632 were significantly downregulated, while the expression of <jats:italic>BCL2<\/jats:italic> was increased compared to the control (<jats:italic>p<\/jats:italic>\u2009&amp;lt;\u20090.05). The results of IVF demonstrated that the treatment of frozen\u2013thawed spermatozoa with 20\u2009\u03bcM Y-27632 increased blastocyst rates compared to the control group (<jats:italic>p<\/jats:italic>\u2009&amp;lt;\u20090.05). In conclusion, the addition of 20\u2009\u03bcM Y-27632 into the freezing extender can improve the functionality and the fertilizing capacity of frozen spermatozoa due to its antioxidative and anti-apoptotic properties.<\/jats:p>","DOI":"10.3389\/fvets.2023.1155048","type":"journal-article","created":{"date-parts":[[2023,7,6]],"date-time":"2023-07-06T18:12:25Z","timestamp":1688667145000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":16,"title":["Improvement of post-thaw quality and fertilizing ability of bull spermatozoa using Rho kinase inhibitor in freezing extender"],"prefix":"10.3389","volume":"10","author":[{"given":"Mina","family":"Behnam","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Reza","family":"Asadpour","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tohid Rezaei","family":"Topraggaleh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hossein","family":"Hamali","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2023,7,6]]},"reference":[{"key":"ref1","doi-asserted-by":"publisher","DOI":"10.5772\/intechope.89789","article-title":"Cryobiology and cryopreservation of sperm. In Cryopreservation-Current Advances and Evaluations","author":"\u00d6zt\u00fcrk","year":"2019","journal-title":"Intech Open"},{"key":"ref2","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1016\/S0378-4320(00)00152-4","article-title":"Basic aspects of frozen storage of semen","volume":"62","author":"Holt","year":"2000","journal-title":"Anim Reprod Sci"},{"key":"ref3","doi-asserted-by":"publisher","first-page":"103","DOI":"10.1016\/j.smallrumres.2006.12.001","article-title":"Protective effect of taurine, glutathione and trehalose on the liquid storage of ram semen","volume":"73","author":"Bucak","year":"2007","journal-title":"Small Rumin Res"},{"key":"ref4","doi-asserted-by":"publisher","first-page":"1682","DOI":"10.1016\/j.theriogenology.2012.06.007","article-title":"The cryobiology of spermatozoa","volume":"78","author":"Benson","year":"2012","journal-title":"Theriogenology"},{"key":"ref5","doi-asserted-by":"publisher","first-page":"655","DOI":"10.1111\/rda.13409","article-title":"Recent advances in bovine sperm cryopreservation techniques with a focus on sperm post-thaw quality optimization","volume":"54","author":"Gr\u00f6tter","year":"2019","journal-title":"Reprod Domest Anim"},{"key":"ref6","doi-asserted-by":"publisher","first-page":"562","DOI":"10.1016\/j.theriogenology.2016.02.007","article-title":"Effects of cryopreservation on sperm viability, synthesis of reactive oxygen species and DNA damage of bovine sperm","volume":"86","author":"G\u00fcrler","year":"2016","journal-title":"Theriogenology"},{"key":"ref7","doi-asserted-by":"publisher","first-page":"878","DOI":"10.1002\/mrd.20686","article-title":"Impact of cryopreservation and reactive oxygen species on DNA integrity, lipid peroxidation, and functional parameters in ram sperm","volume":"74","author":"Peris","year":"2007","journal-title":"Mol Reprod Dev"},{"key":"ref8","doi-asserted-by":"publisher","first-page":"S49","DOI":"10.1016\/j.ijvsm.2017.11.001","article-title":"Evaluation of bull spermatozoa during and after cryopreservation: structural and ultrastructural insights","volume":"6","author":"Khalil","year":"2018","journal-title":"Int J Vet Sci"},{"key":"ref9","doi-asserted-by":"publisher","first-page":"65","DOI":"10.1016\/j.cryobiol.2018.06.001","article-title":"Cryopreservation induces mitochondrial permeability transition in a bovine sperm model","volume":"83","author":"Treulen","year":"2018","journal-title":"Cryobiology"},{"key":"ref10","doi-asserted-by":"publisher","first-page":"354","DOI":"10.1095\/biolreprod66.2.354","article-title":"Sperm apoptosis in fresh and cryopreserved bull semen detected by flow cytometry and its relationship with fertility","volume":"66","author":"Anzar","year":"2002","journal-title":"Biol Reprod"},{"key":"ref11","doi-asserted-by":"publisher","first-page":"456","DOI":"10.1016\/j.rbmo.2010.05.011","article-title":"Implication of apoptosis in sperm cryoinjury","volume":"21","author":"Said","year":"2010","journal-title":"Reprod Biomed Online"},{"key":"ref12","doi-asserted-by":"publisher","first-page":"308","DOI":"10.1017\/S0967199418000254","article-title":"Effects of human sperm cryopreservation on apoptotic markers in normozoospermic and non-normozoospermic patients","volume":"26","author":"Karabulut","year":"2018","journal-title":"Zygote"},{"key":"ref13","doi-asserted-by":"publisher","first-page":"28","DOI":"10.1095\/biolreprod.103.024281","article-title":"Cryopreservation induces an apoptosis-like mechanism in bull sperm","volume":"71","author":"Martin","year":"2004","journal-title":"Biol Reprod"},{"key":"ref14","doi-asserted-by":"publisher","first-page":"7880","DOI":"10.18632\/aging.102295","article-title":"Mechanisms of ROS-induced mitochondria-dependent apoptosis underlying liquid storage of goat spermatozoa","volume":"11","author":"Liu","year":"2019","journal-title":"Aging (Albany NY)"},{"key":"ref15","doi-asserted-by":"publisher","first-page":"677","DOI":"10.18632\/aging.102295","article-title":"Assessment of fresh and frozen-thawed boar semen using an Annexin-V assay: a new method of evaluating sperm membrane integrity","volume":"60","author":"Pe\u00f1a","year":"2003","journal-title":"Theriogenology"},{"key":"ref16","doi-asserted-by":"publisher","first-page":"743","DOI":"10.1016\/S0093-691X(03)00050-5","article-title":"Subtle membrane changes in cryopreserved bull semen in relation with sperm viability, chromatin structure, and field fertility","volume":"60","author":"Januskauskas","year":"2003","journal-title":"Theriogenology"},{"key":"ref17","doi-asserted-by":"publisher","first-page":"1828","DOI":"10.1095\/biolreprod.103.025627","article-title":"Cryopreservation and thawing is associated with varying extent of activation of apoptotic machinery in subsets of ejaculated human spermatozoa","volume":"71","author":"Paasch","year":"2004","journal-title":"Biol Reprod"},{"key":"ref18","doi-asserted-by":"publisher","first-page":"149","DOI":"10.1042\/bj3540149","article-title":"LIM-kinase 2 induces formation of stress fibres, focal adhesions and membrane blebs, dependent on its activation by rho-associated kinase-catalysed phosphorylation at threonine-505","volume":"354","author":"Amano","year":"2001","journal-title":"Biochem J"},{"key":"ref19","doi-asserted-by":"publisher","first-page":"346","DOI":"10.1038\/35070019","article-title":"Caspase-3-mediated cleavage of ROCK I induces MLC phosphorylation and apoptotic membrane blebbing","volume":"3","author":"Sebbagh","year":"2001","journal-title":"Nat Cell Biol"},{"key":"ref20","doi-asserted-by":"publisher","first-page":"446","DOI":"10.1038\/nrm1128","article-title":"Rocks: multifunctional kinases in cell behaviour","volume":"4","author":"Riento","year":"2003","journal-title":"Nat Rev Mol Cell Biol"},{"key":"ref21","doi-asserted-by":"publisher","first-page":"249","DOI":"10.1017\/S0967199406003790","article-title":"Localization of the rho GTPases and some rho effector proteins in the sperm of several mammalian species","volume":"14","author":"Ducummon","year":"2006","journal-title":"Zygote"},{"key":"ref22","doi-asserted-by":"publisher","first-page":"471","DOI":"10.1165\/rcmb.2017-0075OC","article-title":"The rho kinase isoforms ROCK1 and ROCK2 each contribute to the development of experimental pulmonary fibrosis","volume":"58","author":"Knipe","year":"2018","journal-title":"Am J Respir Cell Mol"},{"key":"ref23","doi-asserted-by":"publisher","first-page":"1844","DOI":"10.1093\/humrep\/dep107","article-title":"Mesenchymal stem cells facilitate the derivation of human embryonic stem cells from cryopreserved poor-quality embryos","volume":"24","author":"Cortes","year":"2009","journal-title":"Hum Reprod"},{"key":"ref24","doi-asserted-by":"publisher","first-page":"681","DOI":"10.1038\/nbt1310","article-title":"ROCK inhibitor permits survival of dissociated human embryonic stem cells","volume":"25","author":"Watanabe","year":"2007","journal-title":"Nat Biotechnol"},{"key":"ref25","doi-asserted-by":"publisher","first-page":"96","DOI":"10.1007\/s12015-009-9103-z","article-title":"Presence of a ROCK inhibitor in extracellular matrix supports more undifferentiated growth of feeder-free human embryonic and induced pluripotent stem cells upon passaging","volume":"6","author":"Pakzad","year":"2010","journal-title":"Stem Cell Rev Rep"},{"key":"ref26","doi-asserted-by":"publisher","first-page":"30","DOI":"10.1007\/s11626-011-9471-y","article-title":"Rho kinase inhibitor Y-27632 and Accutase dramatically increase mouse embryonic stem cell derivation","volume":"48","author":"Zhang","year":"2012","journal-title":"In Vitro"},{"key":"ref27","doi-asserted-by":"publisher","first-page":"27","DOI":"10.1016\/j.theriogenology.2019.12.015","article-title":"Optimal concentration of rho-associated coiled-coil kinase (ROCK) inhibitor improved sperm membrane functionality and fertilizing ability of cryopreserved-thawed feline sperm","volume":"144","author":"Tharasanit","year":"2020","journal-title":"Theriogenology"},{"key":"ref28","doi-asserted-by":"publisher","first-page":"8070","DOI":"10.3390\/molecules27228070","article-title":"The effects of different doses of ROCK inhibitor, antifreeze protein III, and boron added to semen extender on semen Freezeability of Ankara bucks","volume":"27","author":"Kara\u015f\u00f6r","year":"2022","journal-title":"Molecules"},{"key":"ref29","doi-asserted-by":"publisher","first-page":"86","DOI":"10.1007\/s12015-009-9107-8","article-title":"Effect of ROCK inhibitor Y-27632 on normal and variant human embryonic stem cells (hESCs) in vitro: its benefits in hESC expansion","volume":"6","author":"Gauthaman","year":"2010","journal-title":"Stem Cell Rev Rep"},{"key":"ref30","doi-asserted-by":"publisher","first-page":"31","DOI":"10.1016\/j.anireprosci.2010.04.005","article-title":"Effects of extender and equilibration time on post-thaw motility and membrane integrity of cryopreserved Gyr bull semen evaluated by CASA and flow cytometry","volume":"120","author":"Leite","year":"2010","journal-title":"Anim Reprod Sci"},{"key":"ref31","doi-asserted-by":"publisher","first-page":"e20210077","DOI":"10.1590\/1984-3143-AR2021-0077","article-title":"The effect of adjusting settings within a computer-assisted sperm analysis (CASA) system on bovine sperm motility and morphology results","volume":"19","author":"O\u2019meara","year":"2022","journal-title":"Anim Reprod"},{"key":"ref32","doi-asserted-by":"publisher","first-page":"9","DOI":"10.3389\/fvets.2022.1043379","article-title":"Fertility-associated biochemical components in seminal plasma and serum of buffalo (Bubalus bubalis) bulls","volume":"9","author":"Almadaly","year":"2022","journal-title":"Front Vet Sci"},{"key":"ref33","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1016\/0378-4320(94)90055-8","article-title":"An osmotic resistance test for bovine semen","volume":"36","author":"Revell","year":"1994","journal-title":"Anim Reprod Sci"},{"key":"ref34","doi-asserted-by":"publisher","first-page":"471","DOI":"10.1530\/REP-09-0094","article-title":"Exression and putative function of fibronectin and its receptor (integrin \u03b15\u03b21) in male and female gametes during bovine fertilization in vitro","volume":"138","author":"Thys","year":"2009","journal-title":"Reproduction"},{"key":"ref35","doi-asserted-by":"publisher","first-page":"20","DOI":"10.1186\/s12610-021-00137-5","article-title":"Effects of vitamin D supplementation in extender on sperm kinematics and apoptosis following the freeze-thaw process in normozoospermic and asthenozoospermic Holstein bulls","volume":"31","author":"Asadpour","year":"2021","journal-title":"Basic Clin Androl"},{"key":"ref36","first-page":"407","article-title":"Determination of aldehydic lipid peroxidation products: malonaldehyde and 4-hydroxynonenal","volume-title":"Methods in Enzymology","author":"Esterbauer","year":""},{"key":"ref37","doi-asserted-by":"publisher","first-page":"777","DOI":"10.1111\/and.12148","article-title":"Effect of cysteine and glutamine added to extender on post-thaw sperm functional parameters of buffalo bull","volume":"46","author":"Topraggaleh","year":"2014","journal-title":"Andrologia"},{"key":"ref38","doi-asserted-by":"publisher","first-page":"193","DOI":"10.1016\/j.anireprosci.2018.09.019","article-title":"Supplementation of extender with coenzyme Q10 improves the function and fertility potential of rooster spermatozoa after cryopreservation","volume":"198","author":"Masoudi","year":"2018","journal-title":"Anim Reprod Sci"},{"key":"ref39","doi-asserted-by":"publisher","first-page":"252","DOI":"10.22074\/IJFS.2021.521116.1057","article-title":"Functional and flow cytometric analysis of buffalo cryopreserved spermatozoa: comparison of different breeds and incubation times","volume":"15","author":"Topraggaleh","year":"2021","journal-title":"Int J Fertil"},{"key":"ref40","doi-asserted-by":"publisher","first-page":"39","DOI":"10.1016\/j.theriogenology.2020.01.056","article-title":"Efficient one-step direct transfer to recipients of thawed bovine embryos cultured in vitro and frozen in chemically defined medium","volume":"146","author":"G\u00f3mez","year":"2020","journal-title":"Theriogenology"},{"key":"ref41","doi-asserted-by":"publisher","first-page":"859","DOI":"10.1016\/0093-691X(95)00271-9","article-title":"Effect of bovine sperm separation by either swim-up or percoll method on success of in vitro fertilization and early embryonic development","volume":"44","author":"Parrish","year":"1995","journal-title":"Theriogenology"},{"key":"ref42","doi-asserted-by":"publisher","first-page":"884","DOI":"10.1071\/rd08103","article-title":"Development and quality of bovine morulae cultured in serum-free medium with specific retinoid receptor agonists","volume":"20","author":"G\u00f3mez","year":"2008","journal-title":"Reprod Fertil Dev"},{"key":"ref43","doi-asserted-by":"publisher","first-page":"683","DOI":"10.1016\/S0093-691X(99)00162-4","article-title":"High bovine blastocyst development in a static in vitro production system using SOFaa medium supplemented with sodium citrate and myo-inositol with or without serum-proteins","volume":"52","author":"Holm","year":"1999","journal-title":"Theriogenology"},{"key":"ref44","doi-asserted-by":"publisher","first-page":"7503","DOI":"10.1038\/s41598-022-11447-7","article-title":"Actions of CSF2 and DKK1 on bovine embryo development and pregnancy outcomes are affected by composition of embryo culture medium","volume":"12","author":"Amaral","year":"2022","journal-title":"Sci Rep"},{"key":"ref45","doi-asserted-by":"publisher","first-page":"191","DOI":"10.1016\/S1472-6483(10)61710-5","article-title":"Vitrification of mammalian spermatozoa in the absence of cryoprotectants: from past practical difficulties to present success","volume":"6","author":"Isachenko","year":"2003","journal-title":"Reprod Biomed Online"},{"key":"ref46","doi-asserted-by":"publisher","first-page":"2781","DOI":"10.3390\/ijms21082781","article-title":"Sperm cryodamage in ruminants: understanding the molecular changes induced by the cryopreservation process to optimize sperm quality","volume":"21","author":"Peris-Frau","year":"2020","journal-title":"Int J Mol Sci"},{"key":"ref47","doi-asserted-by":"publisher","first-page":"195","DOI":"10.3233\/JAD-2007-12210","article-title":"Lipoic acid and N-acetyl cysteine decrease mitochondrial-related oxidative stress in Alzheimer disease patient fibroblasts","volume":"12","author":"Moreira","year":"2007","journal-title":"J Alzheimers Dis"},{"key":"ref48","doi-asserted-by":"publisher","first-page":"1493","DOI":"10.1038\/sj.ki.5002568","article-title":"The role of mitochondria in oxidative and nitrosative stress during ischemia\/reperfusion in the rat kidney","volume":"72","author":"Plotnikov","year":"2007","journal-title":"Kidney Int"},{"key":"ref49","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1016\/j.theriogenology.2014.02.020","article-title":"ROCK inhibitor Y-27632 prevents porcine oocyte maturation","volume":"82","author":"Zhang","year":"2014","journal-title":"Theriogenology"},{"key":"ref50","doi-asserted-by":"publisher","first-page":"1922","DOI":"10.1093\/humrep\/16.9.1922","article-title":"Differential production of reactive oxygen species by subsets of human spermatozoa at different stages of maturation","volume":"16","author":"Gil-Guzman","year":"2001","journal-title":"Hum Reprod"},{"key":"ref51","doi-asserted-by":"publisher","first-page":"498","DOI":"10.1002\/mrd.1115","article-title":"Cryopreservation alters membrane sulfhydryl status of bull spermatozoa: protection by oxidized glutathione","volume":"60","author":"Chatterjee","year":"2001","journal-title":"Mol Reprod Dev"},{"key":"ref52","doi-asserted-by":"publisher","first-page":"675.e13","DOI":"10.1016\/j.urology.2013.11.032","article-title":"Rho-kinase levels in testicular ischemia-reperfusion injury and effects of its inhibitor, Y-27632, on oxidative stress, spermatogenesis, and apoptosis","volume":"83","author":"Cayan","year":"2014","journal-title":"Urology"},{"key":"ref53","first-page":"322","article-title":"Rho\/ROCK signaling pathway and anti-cryodamage ability of human sperm","volume":"25","author":"Tian","year":"2019","journal-title":"Int J Androl"},{"key":"ref54","doi-asserted-by":"publisher","first-page":"221","DOI":"10.1530\/REP-13-0191","article-title":"Phosphorylated AKT preserves stallion sperm viability and motility by inhibiting caspases 3 and 7","volume":"148","author":"Bolanos","year":"2014","journal-title":"Reprod"},{"key":"ref55","doi-asserted-by":"publisher","first-page":"106884","DOI":"10.1016\/j.anireprosci.2021.106884","article-title":"\u201cFine feathers make fine birds\u201d\u2013the mammalian sperm plasma membrane lipid composition and effects on assisted reproduction","volume":"246","author":"Gautier","year":"2022","journal-title":"Anim Reprod Sci"},{"key":"ref56","doi-asserted-by":"publisher","first-page":"578","DOI":"10.4103\/aja.aja_2_20","article-title":"Relationship of sperm plasma membrane and acrosomal integrities with sperm morphometry in Bos taurus","volume":"22","author":"Palac\u00edn","year":"2020","journal-title":"Asian J Androl"},{"key":"ref57","doi-asserted-by":"publisher","first-page":"95","DOI":"10.1016\/j.fertnstert.2004.06.056","article-title":"Impact of sperm morphology on DNA damage caused by oxidative stress induced by \u03b2-nicotinamide adenine dinucleotide phosphate","volume":"83","author":"Said","year":"2005","journal-title":"Fertil Steril"},{"key":"ref58","doi-asserted-by":"publisher","first-page":"65","DOI":"10.3389\/fcell.2018.00065","article-title":"Apoptosis is a demanding selective tool during the development of fetal male germ cells","volume":"6","author":"Bejarano","year":"2018","journal-title":"Front Cell Dev Biol"},{"key":"ref59","doi-asserted-by":"publisher","first-page":"2207","DOI":"10.4049\/jimmunol.148.7.2207","article-title":"Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages","volume":"148","author":"Fadok","year":"1992","journal-title":"J Immunol"},{"key":"ref60","doi-asserted-by":"publisher","first-page":"61","DOI":"10.1007\/s00005-007-0009-7","article-title":"Rho kinase in the regulation of cell death and survival","volume":"55","author":"Shi","year":"2007","journal-title":"Arch Immunol Ther Exp"},{"key":"ref61","doi-asserted-by":"publisher","first-page":"818","DOI":"10.1002\/mrd.20809","article-title":"Efficient propagation of single cells Accutase-dissociated human embryonic stem cells","volume":"75","author":"Bajpai","year":"2008","journal-title":"Mol Reprod Dev"},{"key":"ref62","doi-asserted-by":"publisher","first-page":"1061","DOI":"10.1095\/biolreprod66.4.1061","article-title":"Nature of DNA damage in ejaculated human spermatozoa and the possible involvement of apoptosis","volume":"66","author":"Sakkas","year":"2002","journal-title":"Biol Reprod"},{"key":"ref63","doi-asserted-by":"publisher","first-page":"719","DOI":"10.1111\/j.1745-7270.2005.00108.x","article-title":"Caspase family proteases and apoptosis","volume":"37","author":"Fan","year":"2005","journal-title":"Acta Biochim Biophys Sin"},{"key":"ref64","doi-asserted-by":"publisher","first-page":"451","DOI":"10.1089\/bio.2018.0003","article-title":"ROCK inhibitor during hypothermic storage improves re-expansion rate and quality of goat blastocysts","volume":"16","author":"Saberi","year":"2018","journal-title":"Biopreserv Biobank"},{"key":"ref65","doi-asserted-by":"publisher","first-page":"26","DOI":"10.1095\/biolreprod.113.109769","article-title":"Rescue of vitrified warmed bovine oocytes with rho-associated coiled-coil kinase inhibitor","volume":"89","author":"Hwang","year":"2013","journal-title":"Biol Reprod"},{"key":"ref66","doi-asserted-by":"publisher","first-page":"511","DOI":"10.1262\/jrd.2017-004","article-title":"Effects of vitrification and a rho-associated coiled-coil containing protein kinase 1 inhibitor on the meiotic and developmental competence of feline oocytes","volume":"63","author":"Arayatham","year":"2017","journal-title":"J Reprod Dev"},{"key":"ref67","doi-asserted-by":"publisher","first-page":"159","DOI":"10.1016\/j.fertnstert.2008.09.038","article-title":"Effects of cryopreservation on human sperm deoxyribonucleic acid integrity","volume":"93","author":"Zribi","year":"2010","journal-title":"Fertil Steril"},{"key":"ref68","doi-asserted-by":"publisher","first-page":"8","DOI":"10.1016\/j.ejogrb.2019.10.005","article-title":"The role of sperm DNA fragmentation testing in predicting intra-uterine insemination outcome: a systematic review and meta-analysis","volume":"244","author":"Sugihara","year":"2020","journal-title":"Eur J Obstet Gynecol"},{"key":"ref69","doi-asserted-by":"publisher","first-page":"331","DOI":"10.1093\/humupd\/dmg027","article-title":"Role of sperm chromatin abnormalities and DNA damage in male infertility","volume":"9","author":"Agarwal","year":"2003","journal-title":"Hum Reprod Update"}],"container-title":["Frontiers in Veterinary Science"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2023.1155048\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,7,6]],"date-time":"2023-07-06T18:12:34Z","timestamp":1688667154000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fvets.2023.1155048\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,7,6]]},"references-count":69,"alternative-id":["10.3389\/fvets.2023.1155048"],"URL":"https:\/\/doi.org\/10.3389\/fvets.2023.1155048","relation":{},"ISSN":["2297-1769"],"issn-type":[{"value":"2297-1769","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,7,6]]},"article-number":"1155048"}}