Cryobiology最新文献

{"title":"Pressure-activated disassembly of cryoprotectant supramolecules in isochoric freezing.","authors":"Boris Rubinsky","doi":"10.1016/j.cryobiol.2026.105642","DOIUrl":"https://doi.org/10.1016/j.cryobiol.2026.105642","url":null,"abstract":"<p><p>The efficacy of cryopreservation is fundamentally limited by the difficulty of achieving high intracellular concentrations of cryoprotective agents (CPAs) without inducing osmotic injury or chemical toxicity during loading. This Short Communication advances a thermodynamic hypothesis proposing that intracellular cryoprotection could be achieved through the in situ generation of cryoprotective solutes via pressure-activated disassembly of supramolecular complexes composed of cryoprotectant monomers or oligomers. The physical trigger for this disassembly is the hydrostatic pressure that arises intrinsically during isochoric (constant-volume) freezing: as ice forms, the fixed-volume constraint produces a substantial pressure increase. We propose that the stability of these assemblies is governed by the Helmholtz free energy, and that isochoric pressure shifts the free-energy landscape to favor the dissociated state for assemblies with a negative reaction molar volume. Because the pressures generated during isochoric freezing reach levels known to destabilize supramolecular complexes, this mechanism offers a plausible route for generating CPAs precisely during the freezing process. This approach would decouple cryoprotectant availability from membrane transport and synchronize protection with the onset of freezing. The purpose of this contribution is to articulate the thermodynamic basis and physical plausibility of this mechanism and to motivate future investigation of pressure-mediated preservation strategies.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"123 ","pages":"105642"},"PeriodicalIF":2.1,"publicationDate":"2026-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147855649","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":"Coenzyme Q10 improves the cryopreservation and fertilizing capacity of dromedary camel epididymal sperm.","authors":"Karim A Yaqout, Abdallah M Shahat, Mohamed Hedia, Mostafa M Abou-Ahmed, Abdelraouf M Ghallab","doi":"10.1016/j.cryobiol.2026.105633","DOIUrl":"https://doi.org/10.1016/j.cryobiol.2026.105633","url":null,"abstract":"<p><p>Cryopreservation of epididymal spermatozoa, recovered post-mortem, enables genetic preservation in dromedary camels; however, freeze-thaw procedures induce oxidative stress that hinders spermatozoal function. This study evaluated the effects of supplementing a commercial semen extender (INRA 96) with various concentrations of Coenzyme Q10 (CoQ10; 0, 0.25, 1.00, and 2.00 μM) on post-thaw quality and fertilizing capacity of dromedary camel epididymal spermatozoa. Post-thaw assessments included motility, vitality, morphology, plasma membrane functionality, and acrosome integrity. Oxidative stress markers [total antioxidant capacity (TAC) and malondialdehyde (MDA)] were measured in the extender post-thaw. Cleavage and blastocyst rates after in vitro embryo production (IVEP) were also recorded. Supplementation of the extender with 0.25 μM CoQ10 significantly improved post-thaw motility (48.33% ± 1.67 vs. control 38.33% ± 1.67), vitality (53.33% ± 2.73 vs. control 41.00% ± 2.08), membrane functionality (50.33% ± 4.10 vs. control 47.33% ± 4.26), and acrosome integrity (63.33% ± 2.73 vs. control 46.67% ± 2.40). Additionally, CoQ10 at 0.25 μM enhanced cleavage (30.39% ± 1.86) and blastocyst formation rates (20.59% ± 2.03) compared to the control groups (23.73% ± 1.20 and 15.25% ± 1.45, respectively). Biochemically, 0.25 μM CoQ10 elevated TAC (0.76 ± 0.02 mM/L) and reduced MDA (6.88 ± 0.38 nmol/mL) compared to controls (0.40 ± 0.04 mM/L and 8.99 ± 0.31 nmol/mL, respectively). In contrast, higher CoQ10 concentrations (1.00 and 2.00 μM) impaired both sperm quality and embryo development potential. In conclusion, low-dose CoQ10 supplementation (0.25 μM) showed beneficial outcomes, while concentrations exceeding 1.00 μM were deleterious. Future research should investigate the molecular mechanisms by which CoQ10 protects spermatozoa, including its effects on mitochondrial function and specific antioxidant pathway activation during the cryopreservation process.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"123 ","pages":"105633"},"PeriodicalIF":2.1,"publicationDate":"2026-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147856126","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":"Cryoprotective effects of mangiferin-loaded chitosan nanoparticles on buffalo sperm: Antioxidant, apoptotic, and motility insights.","authors":"Ali Ali El-Raghi, Ekramy M Elmorsy, Shaza A Alyamani, Badriah A Hifni, Ayat B Al-Ghafari, Huda A Al Doghaither, Walaa M Essawi, Aya Aly Elzeer","doi":"10.1016/j.cryobiol.2026.105634","DOIUrl":"https://doi.org/10.1016/j.cryobiol.2026.105634","url":null,"abstract":"<p><p>This study is the first to systematically evaluate the effects of mangiferin-loaded chitosan nanoparticles (MGF-CHNPs) as an additive in a semen extender on the structural and functional characteristics of cryopreserved buffalo sperm, including sperm quality, acrosome integrity, kinematics, antioxidant capacity, apoptosis, ultrastructure, and fertilizing potential. Semen from six healthy, fertile buffalo bulls was cryopreserved using a Tris-based extender supplemented with 0, 25, 50, or 100 μg/mL MGF-CHNPs. Supplementation with 100 μg/mL MGF-CHNPs markedly improved sperm progressive motility, viability, plasma membrane integrity, acrosome reaction, and overall kinematic parameters. This treatment also enhanced total antioxidant capacity and the activities of catalase, superoxide dismutase, and glutathione peroxidase, while significantly reducing oxidative stress markers, including malondialdehyde, hydrogen peroxide, nitric oxide, and nuclear factor kappa B levels (p < 0.05) in the seminal plasma. MGF-CHNPs significantly increased the proportion of viable sperm and decreased apoptotic cells, with upregulation of the anti-apoptotic Bcl-2 gene and downregulation of pro-apoptotic genes, showing the most pronounced effects at 100 μg/mL. Transmission electron microscopy confirmed preservation of the acrosome, plasma membrane, and overall sperm ultrastructure. Importantly, the pregnancy rate of buffalo cows inseminated with semen containing 100 μg/mL MGF-CHNPs increased by 30.66% compared to the control group. In conclusion, adding 100 μg/mL MGF-CHNPs to the freezing extender significantly improves post-thaw sperm quality, antioxidant defenses, structural integrity, and fertility in buffalo sperm.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"123 ","pages":"105634"},"PeriodicalIF":2.1,"publicationDate":"2026-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147834694","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":"Euterpe edulis seed recalcitrance: difficult, yes, but not impossible to genebank.","authors":"Neusa Steiner, Daniela Goeten, Devon Gordon, Miguel P Guerra, Christina Walters","doi":"10.1016/j.cryobiol.2026.105632","DOIUrl":"https://doi.org/10.1016/j.cryobiol.2026.105632","url":null,"abstract":"<p><p>Euterpe edulis is a key palm species in the Atlantic Forest that is threatened by predatory exploitation, habitat reduction and slow seedling recruitment in the wild. The dire need for ex situ conservation is further limited by high sensitivity of embryos to desiccation. In vitro technologies are needed to genebank genetic resources from wild populations using cryogenic storage. We quantified the sensitivity of isolated embryos to desiccation, cryopreservation, and cryoprotectant solutions. We explored cell cryoprotection from lethal ice by balancing water and freezing stress with and without the addition of exogenous cryoprotectants. Embryos were recovered in vitro in MS medium with hormones and antioxidants to achieve normal seedling growth. Viability and germination speed index gradually decreased as embryos were progressively dried. Germination declined from 100% to 42 and 0% in embryos dried to 0.2 gH₂O/gDW, and 0.1 gH₂O/gDW, before cryo-exposure. Nearly 38% of embryos dried to 0.2 gH₂O/gDW survived cryo-exposure, but only 14% developed normal seedlings having both shoot and root development. Cryoprotectant solutions (PVS2 and PVS3) were toxic, reducing germination to about 29% without liquid nitrogen exposure. The work shows that in vitro embryo germination is a feasible approach to produce seedlings quickly and assay embryo survival after water and freezing stresses. The strategy to balance water and freezing stresses proves reliable for E. edulis embryo cryopreservation; however, higher sensitivity to desiccation and unusual water freezing patterns, compared to tissues of other recalcitrant species, explain low survival reported here.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"123 ","pages":"105632"},"PeriodicalIF":2.1,"publicationDate":"2026-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147834818","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 Aurelia aurita larvae, a new model for understanding cryobiology in high-water content marine species.","authors":"Alba Lago, Jesus Troncoso, Estefania Paredes","doi":"10.1016/j.cryobiol.2026.105628","DOIUrl":"https://doi.org/10.1016/j.cryobiol.2026.105628","url":null,"abstract":"<p><p>Cryopreservation of marine species is a key tool for biodiversity conservation, population management, and the advance of aquaculture and biological research. Currently exist significant progress in the cryopreservation of gametes, embryos, larvae and even juveniles of several genus of marine invertebrates. However, cnidarians, and particularly jellyfish, remain largely underexplored. In this study, we report on the first successful cryopreservation of Aurelia aurita ephyrae, the first larval stage of this jellyfish species. This species is characterized by extremely high-water content (96.3 %), which represents a unique challenge in the method of cryopreservation. In this study we done toxicity tests using different concentrations ranging from 0.5 to 6 M of dimethyl sulfoxide (Me₂SO). On the other hand, we designed two cocktails of cryoprotectants and evaluated the efficiency in a protocol of cryopreservation. The first one is based on 1 M Me₂SO + 0.5 M dimethylformamide (DMF) and the second used 1.5 M Me₂SO with 0.04 M trehalose (TRE) and 1 % BSA as a post-thaw supplement. The first protocol achieved 100 % survival immediately after thawing and 33 % survival at 48 h, while the second maintained a consistent 33 % survival across both time points. The damage and viability of the larvae was assessed using live/dead fluorescence dye and allowed confirming the integrity of surviving individuals. These results mark the first demonstration of successful cryopreservation protocol for jellyfish larvae and represent a significant advance for the ex-situ conservation of gelatinous zooplankton. This work lays the foundation for further research into cryopreservation protocols for marine species, understand most better water flow for organisms with high water content and opens new possibilities for marine biodiversity conservation and cnidarian model systems.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"123 ","pages":"105628"},"PeriodicalIF":2.1,"publicationDate":"2026-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147615819","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":"Impact of cryopreservation on pollen viability and morphological integrity of five Indian Crinum species","authors":"Harshid Pulparambil ,&nbsp;P.E. Rajasekharan ,&nbsp;N.S. Pradeep","doi":"10.1016/j.cryobiol.2026.105586","DOIUrl":"10.1016/j.cryobiol.2026.105586","url":null,"abstract":"<div><div><em>Crinum</em> species are ornamental plants of high horticultural value and medicinal importance, known for their bioactive alkaloids with neurological and anticancer properties. Sustainable use and genetic conservation of <em>Crinum</em> require reliable long-term germplasm conservation strategies, and pollen cryopreservation offers an effective approach by enabling year-round breeding and conservation independent of seasonal and geographic constraints. This study presents the first species-comparative evaluation of pollen germination behaviour and cryopreservation responses in five ecologically contrasting Indian <em>Crinum</em> species (<em>C. asiaticum</em>, <em>C. latifolium, C. malabaricum</em>, <em>C. lorifolium and C. viviparum</em>). <em>In vitro</em> pollen germination treatments with varying sucrose (1–10 %) and polyethylene glycol (PEG) concentrations were optimized, species-specific critical moisture contents were determined, and pollen viability was assessed before and after cryostorage at −196 °C for up to 180 days. Distinct, habitat-linked germination responses were observed: aquatic and semi-aquatic species achieved optimal germination (&gt;91 %) only under PEG-mediated osmotic regulation, whereas terrestrial species performed best in sucrose-only treatments. Precise moisture content ranges (12–18 %) were identified as critical for maximizing post-cryopreservation viability, with small deviations resulting in marked germination loss. Cryopreserved pollen retained high functional competence, with minimal viability decline (&lt;5 %) after 180 days, and scanning electron microscopy revealed only minor, non-critical ultrastructural changes. By linking pollen osmotic sensitivity, ecological adaptation, and cryotolerance within a single genus, this study advances pollen cryobiology beyond empirical optimization and provides biologically informed, species-specific protocols for pollen cryobanking. These findings strengthen the scientific basis for <em>ex situ</em> conservation, germplasm exchange, and breeding programmes targeting horticultural and medicinal improvement of <em>Crinum</em> species.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105586"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146073865","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":"Enhanced prevention of cell death by hypothermic storage with propyl gallate","authors":"Xiaodong Li ,&nbsp;Ryan Chan ,&nbsp;Xinglan Li ,&nbsp;Qiunong Guan ,&nbsp;Jingyi Li ,&nbsp;Xiaowei Li ,&nbsp;Haofeng Li ,&nbsp;Shadan Li ,&nbsp;Caigan Du","doi":"10.1016/j.cryobiol.2025.105562","DOIUrl":"10.1016/j.cryobiol.2025.105562","url":null,"abstract":"<div><div>Current hypothermic preservation solutions <strong>are</strong> insufficient to prevent donor organ injury <strong>during hypothermic</strong> storage, which negatively impacts graft functional recovery and survival after transplantation. This study was to evaluate whether supplementation of the preservation solutions with antioxidant propyl gallates (PG) reduced cell injury during hypothermic storage and subsequently rewarming-reoxygenation or transplantation. Human cells and rat aorta were stored in University of Wisconsin (UW) or hyperbranched polyglycerol (HPG) solution supplemented with PG at 4 °C, followed by rewarming-reoxygenation with atmospheric O₂ at 37 °C and isotransplantation, respectively. Cell or tissue injury was measured by lactate dehydrogenase release, flow cytometric and histologic analyses. Here, we showed that PG supplementation significantly increased antioxidative activities of both UW and HPG solutions, and its concentrations for maximum protection of different human cells from hypothermic storage-induced cell death were estimated between 25 and 50 μM, which was also indicated by increased cell survival or decreased cell apoptosis during rewarming-reoxygenation. The PG cytoprotection was correlated with its inhibition of lipid peroxidation, prevention of mitochondrial dysfunction or an increase in ATP content. The enhanced cytoprotection of preservation solutions by PG supplementation was confirmed in hypothermic storage of rat aortas, indicated by less tissue damage and higher levels of tissue ATP during the hypothermic storage and less chronic injury after transplantation. In c<strong>onclusion, cell injury during hypothermic</strong> storage and subsequently rewarming-reoxygenation was substantially prevented by the hypothermic preservation with PG, suggesting that PG is a promising antioxidant for hypothermic preservation of donor cells or organs in transplantation.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105562"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145615983","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":"The effect of whole-body cryostimulation treatments on the profile of selected adipokines and sirtuins in middle-aged individuals with varying degrees of body fatness","authors":"Adrianna Dzidek ,&nbsp;Olga Czerwińska-Ledwig ,&nbsp;Roxana Zuziak ,&nbsp;Joanna Kryst ,&nbsp;Marta Morawiec ,&nbsp;Zbigniew Szyguła ,&nbsp;Tomasz Pałka ,&nbsp;Anna Kurkiewicz-Piotrowska","doi":"10.1016/j.cryobiol.2026.105599","DOIUrl":"10.1016/j.cryobiol.2026.105599","url":null,"abstract":"<div><div>Whole-body cryostimulation (WBC), a widely utilized intervention involving exposure to extremely low temperatures, elicits a range of physiological responses. Growing evidence suggests that WBC may serve as a complementary, non-pharmacological strategy for obesity management and the modulation of metabolic homeostasis. The aim of the study was to evaluate the effects of a series of WBC sessions on the profile of selected adipokines, sirtuins, and carbohydrate metabolism in individuals with normal and elevated BMI. A total of 23 volunteers participated in the study: 12 in the study group (SG; BMI &gt;27) and 11 in the control group (CG; BMI 18.5-25). Blood samples were collected: prior to the 1st (I), after the 9th (II), after the 19th (III) WBC treatment, and two weeks following the cessation of the intervention (IV). The WBC series had a positive impact on fasting blood glucose (I vs II <em>p=</em>0.035; I vs III <em>p=</em>0.039), glycated hemoglobin (I vs IV <em>p=</em>0.004), and asprosin (I vs IV <em>p</em> = 0.020). A positive effect was observed in SG on levels of: asprosin (I vs IV <em>p &lt;</em> 0.001), SIRT1 (<em>p &lt;</em> 0.001), insulin (I vs III <em>p=</em>0.041, I vs IV <em>p=</em>0.013), and HOMA-IR (I vs III <em>p=</em>0.009, I vs IV <em>p=</em>0.009). The WBC series lowered leptin concentrations in SG (I vs III <em>p=</em>0.013) and CG (I vs III <em>p=</em>0.005). However, the downward trend did not continue in SG after the intervention was discontinued. A series of WBC sessions improved carbohydrate metabolism and modulated adipokine and sirtuin profiles, with the most pronounced effects observed in individuals with elevated BMI. However, this effect generally may require a longer treatment series.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105599"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147316791","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":"The post-thaw quality, antioxidant activity, and in vivo fertility of Zaraibi buck semen frozen-stored in the presence of different concentrations of either quercetin or L-arginine","authors":"Essam A. Almadaly ,&nbsp;Mohamed A. Elbendary ,&nbsp;Samia M. Abd El-Rheem ,&nbsp;Ahmed M. Shehabeldin ,&nbsp;Wael B. El-Domany ,&nbsp;Adel A. Ramoun","doi":"10.1016/j.cryobiol.2025.105580","DOIUrl":"10.1016/j.cryobiol.2025.105580","url":null,"abstract":"<div><div>Over the last decade, there has been a growing interest in incorporating specific additives into buck semen extenders to enhance their semen quality, as their antioxidant defense against oxidative stress and lipid peroxidation is insufficient. Due to their strong antioxidant capabilities, quercetin (QUE) and L-arginine (LA) have garnered considerable interest among these additives. Therefore, this study investigates the effect of adding various concentrations of either QUE or LA in the cryopreservation extender on the post-thaw sperm characteristics, kinematics, enzymatic antioxidant activity, and <em>in vivo</em> fertility of buck semen. Ejaculates were collected from 9 healthy <em>Zaraibi</em> bucks using an electroejaculator once a week. Good-quality ejaculates were pooled and dispensed into 7 aliquots; each aliquot was diluted with Tris-egg yolk citrate extender containing: 1) 10 μM QUE; 2) 15 μM QUE; 3) 30 μM QUE; 4) 50 μM QUE; 5) 2 mM LA; 6) 4 mM LA; and 7) the last aliquot was not supplemented with any additive and set as a control (CTRL). Diluted semen samples were equilibrated at 4 °C for 4 h, loaded into French mini straws, sealed, and frozen-stored in liquid nitrogen. Frozen straws were thawed and examined for sperm characteristics and kinematics; also, enzymatic antioxidants, including catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPX), and total antioxidant capacity (TAC), as well as the lipid peroxidation marker malondialdehyde (MDA), were determined. A total of 210 (30 female/group) mature and healthy doe-goats were selected, exposed to estrus synchronization, and inseminated by the prepared frozen-thawed straws to calculate their <em>in vivo</em> fertility rates. The obtained findings revealed that the 30 μM QUE, 50 μM QUE, and 4 mM LA groups had the highest proportions (P &lt; 0.05) of all post-thaw sperm characteristics. Only the 30 μM QUE group had the greatest (P &lt; 0.05) values of all sperm kinematics. Frozen-thawed buck semen's enzymatic antioxidant activity was markedly enhanced by adding either 30 μM QUE or 4 mM LA into the semen extender. The <em>in vivo</em> fertility rates of frozen-thawed straws enriched with either 30 μM QUE (73.33 %) or 4 mM LA (70.00 %) were higher than those of other treatments and the control group (43.33 %). In conclusion, adding either 30 μM QUE or 4 mM LA to the buck semen cryopreservation extender is recommended to improve its post-thaw sperm quality, antioxidant activity, and <em>in vivo</em> fertility.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105580"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145910919","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":"Influence of storage conditions on viability of hematopoietic stem cells and leukocyte subpopulations in fresh and cryopreserved umbilical cord blood samples","authors":"Vladimira Rimac ,&nbsp;Sanja Mazić ,&nbsp;Ines Bojanić","doi":"10.1016/j.cryobiol.2025.105576","DOIUrl":"10.1016/j.cryobiol.2025.105576","url":null,"abstract":"<div><div>Umbilical cord blood (UCB) units are cryopreserved for long-term storage, but it is still not fully investigated how cryopreservation and freezing affect the viability of different cell types. This prospective study evaluated the stability of fresh and cryopreserved UCB samples in various storage conditions using the 7-AAD/annexin V method. UCBs were collected “ex-utero” and processed according to the institutional standard operating procedures. In the stability study, fresh UCB buffy coat (BC) samples were stored at +4 °C and room temperature (RT) for up to 24 h, while thawed samples for up to one and 2 h. After a defined time period, cells were labelled again and analysed. Early apoptosis was most prevalent in CD34⁺ cells and least in T lymphocytes in both fresh and thawed samples. The highest post-thaw recovery was observed for T lymphocytes. The total CD19⁺ (P = 0.001) and CD16+/56+ (P = 0.004) cell counts were statistically significantly reduced when fresh samples were stored at RT. Total NK cell counts were also reduced when samples were stored at +4 °C (P = 0.036). In cryopreserved samples, there was statistically significant differences in total cell counts for all cell populations when samples were stored at RT, and under these storage conditions, early apoptosis of B and NK cells also occurred. The results showed different post-thaw recoveries of leukocyte subpopulations in the samples of cryopreserved UCB units. Cell exposure to the cryoprotective solution post-thaw does not affect total cell count or further development of apoptosis when UCB samples are stored at +4 °C for up to 2 h.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105576"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145862249","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}