Cryobiology最新文献

{"title":"Evaluating the coral microbiome during cryopreservation","authors":"Tori Jefferson ,&nbsp;E. Michael Henley ,&nbsp;Patrick M. Erwin ,&nbsp;Claire Lager ,&nbsp;Riley Perry ,&nbsp;Darya Chernikhova ,&nbsp;Matthew J. Powell-Palm ,&nbsp;Blake Ushijima ,&nbsp;Mary Hagedorn","doi":"10.1016/j.cryobiol.2024.104960","DOIUrl":"10.1016/j.cryobiol.2024.104960","url":null,"abstract":"<div><p>Coral reefs are threatened by various local and global stressors, including elevated ocean temperatures due to anthropogenic climate change. Coral cryopreservation could help secure the diversity of threatened corals. Recently, isochoric vitrification was used to demonstrate that coral fragments lived to 24 hr post-thaw; however, in this study, they were stressed post-thaw. The microbial portion of the coral holobiont has been shown to affect host fitness and the impact of cryopreservation treatment on coral microbiomes is unknown. Therefore, we examined the coral-associated bacterial communities pre- and post-cryopreservation treatments, with a view towards informing potential future stress reduction strategies. We characterized the microbiome of the Hawaiian finger coral, <em>Porites compressa</em> in the wild and at seven steps during the isochoric vitrification process. We observed significant changes in microbiome composition, including: 1) the natural wild microbiomes of <em>P. compressa</em> were dominated by <em>Endozoicomonadace</em>ae (76.5 % relative abundance) and consistent between samples, independent of collection location across Kāneʻohe Bay; 2) <em>Endozoicomonadaceae</em> were reduced to &lt;6.9 % in captivity, and further reduced to &lt;0.5 % relative abundance after isochoric vitrification; and 3) <em>Vibrionaceae</em> dominated communities post-thaw (58.5–74.7 % abundance). Thus, the capture and cryopreservation processes, are implicated as possible causal agents of dysbiosis characterized by the loss of putatively beneficial symbionts (<em>Endozoicomonadaceae</em>) and overgrowth of potential pathogens (<em>Vibrionaceae</em>). Offsetting these changes with probiotic restoration treatments may alleviate cryopreservation stress and improve post-thaw husbandry.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0011224024001159/pdfft?md5=05b9efb5bdb132f9a6521dab92a316cb&pid=1-s2.0-S0011224024001159-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142072278","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ethical assessment of genome resource banking (GRB) in wildlife conservation","authors":"Pierfrancesco Biasetti ,&nbsp;Elena Mercugliano ,&nbsp;Lisa Schrade ,&nbsp;Maria Michela Spiriti ,&nbsp;Frank Göritz ,&nbsp;Susanne Holtze ,&nbsp;Steven Seet ,&nbsp;Cesare Galli ,&nbsp;Jan Stejskal ,&nbsp;Silvia Colleoni ,&nbsp;Daniel Čižmár ,&nbsp;Raffaella Simone ,&nbsp;Thomas B. Hildebrandt ,&nbsp;Barbara de Mori","doi":"10.1016/j.cryobiol.2024.104956","DOIUrl":"10.1016/j.cryobiol.2024.104956","url":null,"abstract":"<div><p>Genome Resources Banks (GRBs) represent vital repositories for the systematic collection, storage, and management of genetic material across various taxa, with a primary objective of safeguarding genetic diversity for research and practical applications. Alongside the development of assisted reproductive techniques (ART), GRBs have evolved into indispensable tools in conservation, offering opportunities for species preservation, mitigating inbreeding risks, and facilitating genetic management across fragmented populations. By preserving genetic information in a suspended state, GRBs serve as backups against population vulnerabilities, potentially aiding in the restoration of endangered species and extending their genetic lifespan. While evidence demonstrates the efficacy of GRBs, ethical considerations surrounding biobanking procedures for wildlife conservation remain largely unexplored. In this article, we will discuss possible ethical issues related to GRBs and the need to ethically monitor biobanking procedures in wildlife conservation. We will then propose a methodological tool, ETHAS, already in use for the ethical self-assessment of assisted reproduction techniques, to assess also biobanking procedures. ETHAS can make it possible to monitor a GRB from its design phase to its actual operation, helping to build biobanking procedures that meet high ethical standards.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0011224024001111/pdfft?md5=443b13411a5e13c20c77748221cd2d9d&pid=1-s2.0-S0011224024001111-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142055197","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Vitrification cryo-foil method for shoot tip cryopreservation and virus eradication in apple","authors":"Min-Rui Wang ,&nbsp;Jun-Hua Bao ,&nbsp;Xiao-Yan Ma ,&nbsp;Zi-Han Yan ,&nbsp;Zhen-Hua Cui ,&nbsp;Li-Ying Zhu ,&nbsp;Dong Zhang ,&nbsp;Qiao-Chun Wang","doi":"10.1016/j.cryobiol.2024.104957","DOIUrl":"10.1016/j.cryobiol.2024.104957","url":null,"abstract":"<div><p>Establishment of a new method for improved shoot tip cryopreservation is crucial to facilitate the long-term preservation of plant germplasm as well as the use of cryotherapy for pathogen eradication. The present study reported a vitrification (V) cryo-foil method for shoot tip cryopreservation and virus eradication in apple. Shoot tip regrowth levels after cryopreservation were comparable among V cryo-foil (53 %), V cryo-plate (46 %) and conventional droplet vitrification (Dr-vi, 48 %). The V cryo-foil is more efficient to perform than Dr-vi as more shoot tips can be cryopreserved by one person. In the histological study applying an image-overlaying strategy, shoot tips cryopreserved by V cryo-foil showed a higher survival chance in the youngest leaf primordia than in the apical dome. When V cryo-foil was tested for virus eradication, fifty-five percent (55 %) of cryo-derived shoots were free of the apple stem pitting virus (ASPV), while none and less than 10 % were free of the apple stem grooving virus (ASGV) and the apple chlorotic leaf spot virus (ACLSV), respectively. Thus, these two viruses were efficiently preserved by V cryo-foil cryopreservation. Noticeably, although the shoot regrowth level was reduced to 27 %, a higher frequency (81 %) of ASPV eradication was achieved when a reduced duration of cryoprotectant exposure was applied in V cryo-foil, supporting the use of insufficient cryoprotection for improved virus eradication.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142046393","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Application of fibrin-based biomaterial for human ovarian tissue encapsulation and cryopreservation as alternative approach for fertility preservation","authors":"Porntip Sirayapiwat ,&nbsp;Christiani A. Amorim ,&nbsp;Wisan Sereepapong ,&nbsp;Punkavee Tuntiviriyapun ,&nbsp;Chanakarn Suebthawinkul ,&nbsp;Paweena Thuwanut","doi":"10.1016/j.cryobiol.2024.104955","DOIUrl":"10.1016/j.cryobiol.2024.104955","url":null,"abstract":"<div><p>This study aimed to investigate the effects of fibrin-based hydrogel encapsulation, with or without vascular endothelial growth factor (VEGF), on follicle quality and cell survival signaling pathways after ovarian tissue cryopreservation. Ovarian cortex donated by seven patients (ages 44–47 years old) was divided into four groups: I) fresh control, II) ovarian tissue without encapsulation (non-FT), III) fibrin (10 mg/mL fibrinogen plus 50 IU/mL thrombin; 10FT) encapsulated tissue without VEGF, and IV) encapsulated tissue with 0.1 μg/mL VEGF (10FT-VEGF), followed by a slow freezing process. Evaluation criteria included normal follicle morphology, density, cell proliferation, apoptosis, and metabolism signaling pathways (<em>BAX/BCL-2</em> ratio, <em>CASPASE-3</em> and <em>9</em>, <em>ATP-6</em> genes, VEGF-A, and ERK-1/2 protein expression levels). Major outcomes revealed that the percentages of morphologically normal follicles and density were significantly decreased by cryopreservation. Ovarian tissue encapsulation using the 10FT formulation (with or without VEGF) could maintain the ERK-signaling cascade, which was comparable to the fresh control. Among the frozen-thawed cohorts, the <em>BAX/BCL-2</em> ratio, <em>CASPASE-3</em>, <em>CASPASE-9</em>, and <em>ATP-6</em> expression levels were unfavorable in the non-FT group. However, statistically different results, including VEGF-A expression levels, were not detected. Collectively, our present data demonstrated the first applicable biomaterial matrix for human ovarian tissue encapsulation which might create an optimal intra-ovarian cortex environment during cryopreservation. Further studies to optimize hydrogel polymerization should be expanded, given the potential benefits for cancer patients who wish to preserve fertility through ovarian tissue cryopreservation.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142139476","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cryopreservation of human teeth using vitrification method with cryoprotectant cocktails and N-acetylcysteine for banking and clinical applications","authors":"Peiru Jiang ,&nbsp;Shan Hu ,&nbsp;Chengxiang Zheng ,&nbsp;Yinzhuo Liu ,&nbsp;Qixuan Zhang ,&nbsp;Lei Dou","doi":"10.1016/j.cryobiol.2024.104959","DOIUrl":"10.1016/j.cryobiol.2024.104959","url":null,"abstract":"<div><p>Preserving freshly-extracted healthy human teeth offers an optional resource for potential tooth transplantation and cell therapy. This study aimed to assess the impact of vitrification, utilizing a blend of cryoprotectant agents and N-acetylcysteine (NAC), on the cryopreservation of periodontal ligament tissues, and investigate the underlying mechanisms of NAC on the tooth cryopreservation. Periodontal ligament cells were isolated from freshly-extracted healthy human permanent teeth, and cell sheets of PDLCs were fabricated. The samples including cell sheets, freshly-extracted human and rat teeth were cryopreserved with or without NAC for three months. The viability, ROS level, gene expressions and microstructure of PDLCs within cell sheets were assessed. The expression of SOD-2, Caspase3, LC3A/B and Catalase were evaluated through western blotting. Histological assessments of cryopreserved cell sheets and teeth were conducted. PDLCs were isolated from cryopreserved teeth, and their immunophenotype and differentiation ability were evaluated. The data was analyzed using one-way analysis of variance. The vitrification method showed good performance in preserving the viability and differentiation potential of PDLCs. Cryopreservation supplemented with NAC improved the survival rate of PDLCs, enhanced osteogenic differentiation ability, upregulated the expression of SOD-2 and Catalase, and inhibited cell apoptosis. Additionally, mRNA sequencing analysis revealed a significant activation of the PI3K-AKT pathway following cryopreservation via vitrification. Adding a PI3K-AKT activator improved the survival rates of PDLCs post-cryopreservation. The vitrification strategy combining various CPAs and NAC proved to be feasible for tooth cryopreservation. Targeting the PI3K-AKT pathway may improve the efficacy of tooth cryopreservation.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-09-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142055196","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring freeze-injury mechanism through ion-specific analysis of leachate from reversibly versus irreversibly injured spinach (Spinacia oleracea L.) leaves","authors":"Rajeev Arora","doi":"10.1016/j.cryobiol.2024.104954","DOIUrl":"10.1016/j.cryobiol.2024.104954","url":null,"abstract":"<div><p>The present study analyzed four cations (K⁺, Ca²⁺, Mg²⁺, Fe²⁺) in leachate from freeze-injured spinach (<em>Spinacia oleracea</em> L. ‘Reflect’) leaves exposed for four freezing-durations (FDs) (0.5, 3.0, 5.5, 10.5 h) at −4.8 °C. Comparison of electrolyte leakage from right-after-thaw with that after 6-d recovery revealed that injury at 0.5 or 3 h FDs was recoverable but irreversible at 5.5 or 10.5 h FDs. Data suggests leakage of K⁺, the most abundant cation in leachate, can serve as a proxy for total electrolyte-leakage in determining plant freezing-tolerance and an ionic marker discerning moderate vs. severe injury. Quantitative correspondence between Ca²⁺- and K⁺-leakage supports earlier proposition that leaked K⁺ induces loss of membrane-Ca²⁺, which, in turn, promotes further K⁺-leakage due to weakened membrane. Reduced/undetectable Fe²⁺ in leachate at longer FDs suggests activation of Fenton reaction converting soluble Fe²⁺ into insoluble Fe³⁺. Enhanced Mg²⁺-leakage at greater freeze-injury suggests structural/functional impairment of chlorophyll/chloroplast complex.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-09-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141995515","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of repeated cryostimulation exposures on sleep and wellness in healthy young adults","authors":"Coralie Arc-Chagnaud ,&nbsp;Olivier Dupuy ,&nbsp;Manuela Garcia ,&nbsp;Laurent Bosquet ,&nbsp;Romain Bouzigon ,&nbsp;Robin Pla","doi":"10.1016/j.cryobiol.2024.104948","DOIUrl":"10.1016/j.cryobiol.2024.104948","url":null,"abstract":"<div><p>The aim of this study was to evaluate the effects of daily whole-body cryostimulation (WBC) sessions during 5 consecutive days on wellness and sleep parameters in healthy young men and women. Twenty healthy subjects (9 women; 11 men) aged 23.1 ± 2.6 years old participated in this randomized protocol, with 5 consecutive days with (CRYO) or without WBC (CONT) exposure. Sleep was analyzed over the 5 nights in each condition. Sleep quality and quantity were assessed via actimetry, cerebral activity and questionnaires. Nocturnal heart rate variability (HRV) was also recorded and questionnaires were given to assess wellness and mood. Repeated WBC exposures had a beneficial impact on mood and anxiety. It also improved subjective sleep quality (scored from 3.6 ± 0.5 pre to 3.9 ± 0.3), especially in women. Also, repeated WBC sessions modulated sleep architecture by increasing slow wave sleep duration (+7.3 ± 16.8 min) during the nights without impacting other sleep parameters, nor nocturnal HRV. In conclusion, repeated WBC seems to be an effective strategy to improve slow wave sleep duration in healthy young subjects. The reported psychological improvements may better benefit women than men.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0011224024001032/pdfft?md5=a8807aa7f34739b065c695da2ce5d7da&pid=1-s2.0-S0011224024001032-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141888734","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Sub-zero non-freezing of vascularized composite allografts in a rodent partial hindlimb model","authors":"I. Filz von Reiterdank ,&nbsp;P. Tawa ,&nbsp;Y. Berkane ,&nbsp;E. de Clermont-Tonnerre ,&nbsp;A.T. Dinicu ,&nbsp;C. Pendexter ,&nbsp;M. Goutard ,&nbsp;A.G. Lellouch ,&nbsp;A.B. Mink van der Molen ,&nbsp;J.H. Coert ,&nbsp;C.L. Cetrulo Jr ,&nbsp;K. Uygun","doi":"10.1016/j.cryobiol.2024.104950","DOIUrl":"10.1016/j.cryobiol.2024.104950","url":null,"abstract":"<div><p>Ischemia is a major limiting factor in Vascularized Composite Allotransplantation (VCA) as irreversible muscular injury can occur after as early as 4–6 h of static cold storage (SCS). Organ preservation technologies have led to the development of storage protocols extending rat liver ex vivo preservation up to 4 days. Development of such a protocol for VCAs has the added challenge of inherent ice nucleating factors of the graft, therefore, this study focused on developing a robust protocol for VCA supercooling. Rodent partial hindlimbs underwent subnormothermic machine perfusion (SNMP) with several loading solutions, followed by a storage solution with cryoprotective agents (CPA) developed for VCAs. Storage occurred in suspended animation for 24h and VCAs were recovered using SNMP with modified Steen. This study shows a robust VCA supercooling preservation protocol in a rodent model. Further optimization is expected to allow for its application in a transplantation model, which would be a breakthrough in the field of VCA preservation.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-08-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141970836","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A practical device to produce consistent, controlled cryoinjury on the skin","authors":"Emilee Herringshaw ,&nbsp;Cemre Turk ,&nbsp;Sandeep Korupolu ,&nbsp;Moriel Tagar Sar-el ,&nbsp;R. Rox Anderson ,&nbsp;Yakir Levin ,&nbsp;Joshua Tam","doi":"10.1016/j.cryobiol.2024.104951","DOIUrl":"10.1016/j.cryobiol.2024.104951","url":null,"abstract":"<div><p>It has long been known that sensitivity to cold-induced damage can vary greatly between different cell types, with lethal temperatures reportedly ranging from −2 °C for canine osteocytes, to −70 °C for mammary adenocarcinomas. This suggests that, for certain applications, “therapeutic windows” may exist wherein the cryosurgery temperature could be controlled to specifically target more cold-sensitive cell types, while sparing less sensitive cells. However, this potential selectivity has not been developed into practical clinical treatments, in part because of a lack of available investigative tools that can provide consistent, reproducible cooling within the desired temperature range. Here we describe an experimental cryosurgery tool that allows user control over the three key cryosurgery parameters – temperature, pressure, and duration. The tool is composed of inexpensive components that are generally accessible in most laboratory settings, and could be a practical investigative tool for developing and optimizing novel topical cryosurgery approaches.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-08-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141916306","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of naringenin on post-thaw quality, fertility-associated gene expression and fertilization potential of buffalo (Bubalus bubalis) bull sperm","authors":"Ghulam Shabbir Khan ,&nbsp;Muhammad Zahid Tahir ,&nbsp;Muhammad Yasir Zahoor ,&nbsp;Hifz-ul-Rahman ,&nbsp;Amjad Riaz","doi":"10.1016/j.cryobiol.2024.104953","DOIUrl":"10.1016/j.cryobiol.2024.104953","url":null,"abstract":"&lt;div&gt;&lt;p&gt;Our objectives were to explore the effect of naringenin addition in the semen extender on the post-thaw 1) sperm quality, 2) fertility-associated gene expression, and 3) fertilization potential of buffalo bull sperm. In experiment 1, semen samples (n = 32) from four Nili-Ravi buffalo bulls were pooled (n = 8) and diluted with the tris-citric acid (TCF-EY) extender containing different concentrations of naringenin, i.e., placebo (DMSO), 0 (control), 50, 100, 150 and 200 μM naringenin. After dilution, semen samples were packed in 0.5 mL French straws, cryopreserved and analyzed for post-thawed sperm quality and gene expression. Computer-assisted Semen Analysis, Hypo-osmotic Swelling test, Normal Apical Ridge assay, Rhodamine 123, Acridine orange, Propidium iodide staining and Thiobarbituric Acid Reactive Substances assay were performed to assess sperm motility parameters, plasma membrane functionality, acrosome integrity, mitochondrial membrane potential, DNA integrity, viability and lipid peroxidation, respectively. Expression levels of sperm acrosome-associated SPACA3, DNA condensation-related PRM1, anti-apoptotic BCL2, pro-apoptotic BAX, and oxidative stress-associated ROMO1 genes were evaluated through qPCR. Results revealed that total and progressive motility, plasma membrane functionality, acrosome integrity, mitochondrial membrane potential, DNA integrity and viability were higher (P &lt; 0.05) with 50, 100 and 150 μM naringenin compared to 200 μM naringenin, placebo and control groups. Moreover, all naringenin-treated groups improved catalase activity, and reduced lipid peroxidation compared to placebo and control groups (P &lt; 0.05). Relative expression levels of SPACA3 and PRM1 genes were higher (P &lt; 0.05) with 150 μM naringenin compared to all groups except 100 μM (P &gt; 0.05). No difference (P &gt; 0.05) in the expression level of BCL2 gene was observed among all groups. Furthermore, BAX gene was expressed higher (P &lt; 0.05) in the 200 μM naringenin group, whereas no difference (P &gt; 0.05) in expression was noticed among the remaining groups. In addition, ROMO1 gene was expressed lower (P &lt; 0.05) in all naringenin-treated groups compared to the control. In experiment 2, the &lt;em&gt;in vivo&lt;/em&gt; fertility of semen doses (n = 400; 200/group) containing optimum concentration of naringenin (150 μM; depicted better &lt;em&gt;in vitro&lt;/em&gt; sperm quality in experiment 1) was compared with control during the breeding season. Buffaloes were inseminated 24 h after the onset of natural estrus and palpated transrectal for pregnancy at least 60 days post-insemination. The fertility rate of 150 μM naringenin group was higher (P = 0.0366) compared to the control [57.00 ± 0.03 % (114/200) &lt;em&gt;vs.&lt;/em&gt; 46.50 ± 0.04 % (93/200), respectively]. Taken together, it is concluded that naringenin supplementation in semen extender improves post-thaw quality, fertility-associated gene expression and fertilization potential of buffalo bull sperm, more ap","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-08-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141981945","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}