CryobiologyPub Date : 2026-03-01Epub Date: 2026-02-05DOI: 10.1016/j.cryobiol.2026.105587
William A. Rowe, Chris Rowe Taitt
{"title":"In Memoriam: Dr. Arthur W. Rowe","authors":"William A. Rowe, Chris Rowe Taitt","doi":"10.1016/j.cryobiol.2026.105587","DOIUrl":"10.1016/j.cryobiol.2026.105587","url":null,"abstract":"","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105587"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146131441","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}
CryobiologyPub Date : 2026-03-01Epub Date: 2025-12-13DOI: 10.1016/j.cryobiol.2025.105568
Merve Deniz Tanrıkulu , Mesut Çevik , Mustafa Numan Bucak , Alper Koçyiğit
{"title":"Extracellular vesicles as emerging bioactive modulators in sperm cryopreservation: Mechanisms and therapeutic potentials","authors":"Merve Deniz Tanrıkulu , Mesut Çevik , Mustafa Numan Bucak , Alper Koçyiğit","doi":"10.1016/j.cryobiol.2025.105568","DOIUrl":"10.1016/j.cryobiol.2025.105568","url":null,"abstract":"<div><div>Sperm cryopreservation is a cornerstone of assisted reproduction and genetic conservation, yet the freeze-thaw process induces substantial structural and functional damage to spermatozoa. Recent progress in the field has positioned extracellular vesicles (EVs) as potential therapeutic tools, primarily because of their capacity to transport functional biomolecules that help counteract the cellular damage caused by cryopreservation. We then examine the types of cellular and molecular damages encountered during sperm cryopreservation and synthesize current evidence on the protective effects of EVs in this context. Particular attention is given to the mechanisms by which EVs may preserve sperm integrity, including membrane stabilization, antioxidant delivery, and modulation of signaling pathways. Despite encouraging findings, challenges remain, including the standardization of EVs isolation methods, optimal dosing, and delivery strategies. We also discuss the need for mechanism-of-action studies and multi-omics approaches to better understand EV-sperm interactions. Finally, future directions are outlined, including EVs engineering and clinical translation. Overall, EVs emerge as promising modulators in cryobiology, with potential to improve sperm preservation outcomes and fertility restoration strategies. This review offers an in-depth examination of EVs, covering the processes involved in their formation, various strategies employed for their isolation, and the techniques used to assess their properties. It also highlights their sources and functional significance in the male reproductive system.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105568"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733112","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}
CryobiologyPub Date : 2026-03-01Epub Date: 2025-12-23DOI: 10.1016/j.cryobiol.2025.105567
Cuiting Yang , Yan Zhang , Chun Wang , Lingxi Zhu , Ming Zhang
{"title":"Cryopreservation decreased lipopolysaccharide-induced immune response of endometrial stromal cells via inhibiting the expression of TRIF","authors":"Cuiting Yang , Yan Zhang , Chun Wang , Lingxi Zhu , Ming Zhang","doi":"10.1016/j.cryobiol.2025.105567","DOIUrl":"10.1016/j.cryobiol.2025.105567","url":null,"abstract":"<div><div>In the study, we investigated the effect of cryopreservation on the immune response of endometrial stromal cells (ESCs) to lipopolysaccharide (LPS). We exposed both frozen-thawed and untreated ESCs to LPS and measured cytokine levels as well as genes associated with the LPS-TLR4-cytokine signaling pathway. Results revealed that LPS induced significant upregulation of <em>IL-1β</em>, <em>IL-6</em>, <em>IL-8</em>, and <em>TNF-α</em> in vitro cultured untreated cells compared to frozen-thawed ones. Furthermore, mRNA levels related to the TRIF pathway, including <em>TRAM</em>, <em>TRAF-6</em>, and <em>IRF-7</em>, <em>NF-κB</em>, and <em>JNK</em> were significantly upregulated in untreated ESCs after LPS-stimulation but not in frozen-thawed cells. Moreover, transcription levels of TRIF signaling-related genes were notably lower in frozen-thawed cells. Additionally, TRIM69 overexpression rescued the cryopreservation-induced suppression of immune response. Overall, our findings suggest that cryopreservation attenuates the LPS-induced immune response in ESCs by inhibiting the TRIF pathway.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105567"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145827118","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}
CryobiologyPub Date : 2026-03-01Epub Date: 2026-01-20DOI: 10.1016/j.cryobiol.2026.105585
Jasmine Bagge , Gustav Hagberg , Anna Ermund , Anna Casselbrant , Edina Sehiç , John Mackay Søfteland , Mats Hellström , Gustaf Herlenius , Mihai Oltean
{"title":"The histologic and molecular alterations during the cold storage of the ascending human colon","authors":"Jasmine Bagge , Gustav Hagberg , Anna Ermund , Anna Casselbrant , Edina Sehiç , John Mackay Søfteland , Mats Hellström , Gustaf Herlenius , Mihai Oltean","doi":"10.1016/j.cryobiol.2026.105585","DOIUrl":"10.1016/j.cryobiol.2026.105585","url":null,"abstract":"<div><div>Inclusion of a colon segment in visceral grafts is increasing as this may improve water and electrolyte absorption and ultimately improve recipients’ renal function. In contrast with the vast knowledge on the ischemia/reperfusion injury in the small bowel, similar data on the colon is almost inexistent. Using light microscopy, immunofluorescence and Western blot, we assessed several histological and molecular changes following the cold storage (CS) of the ascending colon of 18 human brain-dead organ donors for up to 24 h. CS induced progressive yet limited epithelial detachment and goblet cell depletion. Tight junction proteins claudin-1, claudin-3, claudin-4 and occludin were well preserved during the first 14 h of cold storage but its tissue expression decreased following 24 h of CS as evidenced by Western blot and immunofluorescence. ZO-1 expression remained unchanged throughout the 24 h of CS and showed a strong and continuous immunostaining along the entire epithelial lining. Expression of Na+/H+ exchanger (NHE)-1 and 2 remained unaffected by CS. Enterocyte apoptosis increased significantly after 14 h of CS. The current data indicate that colonic mucosa can withstand at least 14 h of cold ischemia without significant histological or molecular changes and implies that the small bowel remains the most vulnerable part of a prospective visceral allograft.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105585"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146017591","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}
CryobiologyPub Date : 2026-03-01Epub Date: 2025-12-30DOI: 10.1016/j.cryobiol.2025.105572
Y. Zhang, K. Beilby, S. Catt, M. Pangestu
{"title":"Optimizing ovarian tissue preparation methods for vitrification: A two-part study on slicing methods and tissue size effects","authors":"Y. Zhang, K. Beilby, S. Catt, M. Pangestu","doi":"10.1016/j.cryobiol.2025.105572","DOIUrl":"10.1016/j.cryobiol.2025.105572","url":null,"abstract":"<div><div>Ovarian tissue cryopreservation by vitrification is well-established, but the effects of procedural differences in the preparatory steps, such as dissection method and tissue size, remain underexplored. This study compared the thickness of cortical tissue pieces and the normality of primordial follicles prepared with either a Stadie Riggs tissue slicer or a conventional scalpel. Part two of the study assessed the viability of follicles vitrified within either 0.5–1 × 5 × 5 mm (thickness × length × width) or 0.5–1 × 10 × 5 mm pieces of cortical tissue. In part one, the proportion of morphologically normal follicles and section thickness did not differ by method (<span><math><mrow><mi>p</mi><mo>=</mo><mn>0</mn><mo>.</mo><mn>309</mn></mrow></math></span> and <span><math><mrow><mi>p</mi><mo>=</mo><mn>0</mn><mo>.</mo><mn>986</mn></mrow></math></span>), although thickness varied by operator (<span><math><mrow><mi>p</mi><mo>=</mo><mn>0</mn><mo>.</mo><mn>028</mn></mrow></math></span>) without an interaction between method and operator. Upon visual H&E analysis, ovarian pieces prepared with the Stadie Riggs devices were more uniform in thickness and the sectioned edge less jagged than those prepared with a conventional scalpel. In part two, follicles were assessed on matched H&E, Ki67 and <span><math><mi>γ</mi></math></span>H2AX stained sections before and after four days of <em>in vitro</em> culture. Fresh tissue processed on day 0 yielded higher proportions of ‘good’ outcomes for morphology, proliferation, and DNA-damage compared to vitrified tissues (<span><math><mrow><mi>p</mi><mo><</mo><mn>0</mn><mo>.</mo><mn>001</mn></mrow></math></span>, <span><math><mrow><mi>p</mi><mo><</mo><mn>0</mn><mo>.</mo><mn>001</mn></mrow></math></span> and <span><math><mrow><mi>p</mi><mo>=</mo><mn>0</mn><mo>.</mo><mn>001</mn></mrow></math></span>, respectively). By day 4, group differences were not significant. Direct comparisons between vitrified 0.5–1 × 5 <span><math><mo>×</mo></math></span> 5 mm and 0.5–1 × 10 <span><math><mo>×</mo></math></span> 5 mm vitrified pieces did not show significant differences at either time point (all <span><math><mrow><mi>p</mi><mo>></mo><mn>0</mn><mo>.</mo><mn>05</mn></mrow></math></span>); pooled analyses yielded odds ratios <span><math><mo>∼</mo></math></span>1 with no heterogeneity. In summary, both preparation methods provide comparable quantitative outcomes, with operator standardization appearing more influential than device choice. For vitrified ovarian cortex, both tissue sizes are interchangeable, and fresh tissue is superior at baseline. Considering the smoother cuts observed, we recommend Stadie Riggs preparation with either 0.5–1 × 5 <span><math><mo>×</mo></math></span> 5 or 0.5–1 × 10 <span><math><mo>×</mo></math></span> 5 mm fragments.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105572"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145877982","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}
CryobiologyPub Date : 2026-03-01Epub Date: 2025-12-30DOI: 10.1016/j.cryobiol.2025.105574
Ali Ali El-Raghi , Amer K. Mohammed , Ekramy M. Elmorsy , Mahmoud A.E. Hassan , Walaa M. Essawi , Suzan Shawky Abuelkasem Mohamed , Abeer M.E. Hassan , Soha A. Hassan
{"title":"Nanophytosomal delivery of resveratrol as an effective strategy for enhancing frozen–thawed bovine sperm quality","authors":"Ali Ali El-Raghi , Amer K. Mohammed , Ekramy M. Elmorsy , Mahmoud A.E. Hassan , Walaa M. Essawi , Suzan Shawky Abuelkasem Mohamed , Abeer M.E. Hassan , Soha A. Hassan","doi":"10.1016/j.cryobiol.2025.105574","DOIUrl":"10.1016/j.cryobiol.2025.105574","url":null,"abstract":"<div><div>The aim of this study was to assess the effects of supplementing semen extender with free resveratrol (FR) or resveratrol-loaded nano-phytosomes (RPNPs) on the quality, antioxidant capacity, enzymatic activities, and apoptotic genes of cryopreserved bovine sperm. Semen samples were collected from five proven fertility Holstein cattle bulls aged 4–6 years using the artificial vagina method. Pooled semen was cryopreserved in a Tris-based extender supplemented with free resveratrol (FR) at a concentration of 90 μg/mL and RPNPs at concentrations of 30 μg/mL (RPNPs30), 60 μg/mL (RPNPs60), and 90 μg/mL (RPNPs90). Cryopreserved semen without any additives served as the control. Supplementing the freezing medium with free resveratrol (FR) or RPNPs at 60 or 90 μg/mL significantly enhanced sperm progressive motility, viability, membrane integrity, and kinematic parameters (p < 0.05). Additionally, RPNPs60 and RPNPs90 treated samples exhibited superior antioxidant activities (TAC, GPX, and SOD), reduced lipid peroxidation, and improved enzymatic activities in seminal plasma compared to the control group (p < 0.05). The supplementation of FR or RPNPs significantly improved the transcription of apoptotic genes such as <em>caspase 3, Bcl2,</em> and <em>Bax</em> in sperm. Electron microscopy observations revealed that fortifying the freezing media with 60 or 90 μg of RPNPs maintained the integrities of the acrosome and plasma membrane, and preserved the ultrastructure integrity of cryopreserved bovine spermatozoa. The docking analysis revealed that the binding affinity with key sperm function biomarkers, such as CAT, SOD, GPX, Caspase, Bax, and BCL2, exhibited binding energies of −8.5, −7.9, −7.5, −10.4, −9.0, and −9.8 kcal/mol, respectively. The addition of 60–90 μg RPNPs to the bovine freezing extender improved post-thawed sperm quality.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105574"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145877962","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}
CryobiologyPub Date : 2026-03-01Epub Date: 2025-12-11DOI: 10.1016/j.cryobiol.2025.105570
Wangjie Sun , Xueqiang Dong , Xian Wang , Pengsong Yuan , Maoqiong Gong
{"title":"Numerical thermal analysis and optimization of vitrification cooling strategies for porcine heart cryopreservation","authors":"Wangjie Sun , Xueqiang Dong , Xian Wang , Pengsong Yuan , Maoqiong Gong","doi":"10.1016/j.cryobiol.2025.105570","DOIUrl":"10.1016/j.cryobiol.2025.105570","url":null,"abstract":"<div><div>Vitrification-based cryopreservation offers a promising approach for long-term organ storage; however, achieving uniform cooling in large-volume organs remains a significant challenge due to the severe thermal gradients that can induce cracking and ice formation within these organs. Using a high-fidelity 3D finite element model of a porcine heart from MRI data, we systematically investigate how key parameters, including initial cooling temperature, glass-phase cooling rate, convective heat transfer coefficient, cryoprotective agent (CPA) thermal conductivity, and container geometry, impact vitrification performance. Our simulation results reveal that an optimized protocol, with an initial cooling temperature of −10 °C, a glass-phase cooling rate of 1.0 °C·min<sup>−1</sup>, and a convective heat transfer coefficient of 250 W∙m<sup>−2</sup>∙°C<sup>−1</sup>, successfully balances cooling efficiency with thermal uniformity. Under these conditions, the average central cooling rates of both heart tissue and CPA were confirmed to exceed 1.4 °C·min<sup>−1</sup>, surpassing the critical cooling rate of VS55 + 0.6 M sucrose (<1.0 °C·min<sup>−1</sup>), while limiting the peak internal temperature difference to below 25.5 °C to minimize thermal stress. Furthermore, we found that enhancing CPA thermal conductivity, reducing its specific heat capacity, and utilizing cylindrical containers significantly improved temperature uniformity and reduced equilibration time. These findings provide critical quantitative insights for optimizing vitrification strategies and establish a robust theoretical foundation for designing safer and more efficient clinical cryopreservation protocols for large organs.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105570"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145733222","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}
CryobiologyPub Date : 2026-03-01Epub Date: 2025-12-22DOI: 10.1016/j.cryobiol.2025.105563
Elham Ashrafi , Janet A.W. Elliott
{"title":"Characterizing the cryobiological response of a mouse cardiac endothelial cell line to interrupted slow cooling (graded freezing)","authors":"Elham Ashrafi , Janet A.W. Elliott","doi":"10.1016/j.cryobiol.2025.105563","DOIUrl":"10.1016/j.cryobiol.2025.105563","url":null,"abstract":"<div><div>Mouse cardiac endothelial cells play a critical role in cardiovascular research and disease; hence, high quality cryopreservation of this cell type is necessary to provide on-demand access to the cells, quality control, and consistent outcomes between different experiments and batches. However, the cryopreservation of cardiac endothelial cells has not been studied as much as other endothelial cells. The objective of this study was to characterize the cryobiological response of an immortalized line of mouse cardiac endothelial cells in suspension using an interrupted slow cooling approach (graded freezing) and to validate cryopreservation protocols that provide high post-thaw viability and function. Three different cryopreservation protocols were validated using different cryoprotectant solutions: <em>i</em>) 10 % dimethyl sulfoxide + 90 % fetal bovine serum, <em>ii</em>) 5 % dimethyl sulfoxide + 6 % hydroxyethyl starch in complete growth medium, and <em>iii</em>) 10 % glycerol in complete growth medium; all three protocols resulted in high post-thaw membrane integrity assessed using dual fluorescent dye Syto13/GelRed. Two functional assays: <em>i</em>) tube forming on Matrigel representing angiogenesis and <em>ii</em>) nitric oxide synthesis using DAF-FM (4-amino-5-methylamino-2′,7′-difluorofluorescein) fluorescent dye revealed that cryopreserved cells retained comparable post-thaw function to non-cryopreserved cells. This study provides practical insight into successful cryopreservation of mouse cardiac endothelial cells.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105563"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145818483","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}
CryobiologyPub Date : 2026-03-01Epub Date: 2026-01-19DOI: 10.1016/j.cryobiol.2026.105583
Zhimin Wang , Chao Tong , Miaomiao Xu , Shanshan Feng , Mengqi Dong , Rongjia Rao , Xianqiang Wang , Wei Feng , Changwei Zhang , Zhan Hu , Li Wang , Shengshou Hu , Bingying Zhou
{"title":"Optimized methods for the functional cryopreservation of adult human primary cardiomyocytes","authors":"Zhimin Wang , Chao Tong , Miaomiao Xu , Shanshan Feng , Mengqi Dong , Rongjia Rao , Xianqiang Wang , Wei Feng , Changwei Zhang , Zhan Hu , Li Wang , Shengshou Hu , Bingying Zhou","doi":"10.1016/j.cryobiol.2026.105583","DOIUrl":"10.1016/j.cryobiol.2026.105583","url":null,"abstract":"<div><div>Adult human primary cardiomyocytes (hPCMs) is a high-fidelity and informative cardiac model that is expected to advance our knowledge of the human heart. However, currently, no method exists that recovers hPCMs from cryopreservation with high efficiency, limiting their use as a versatile research model. Based on our previous success with isolated hPCMs, we designed a new strategy that cryopreserves myocardial tissue at a specific step during hPCM dissociation, i.e., after sectioning tissue chunks into tissue slices. This method yielded cell viabilities comparable to that of freshly isolated hPCMs, and surpassed the performance of cell isolation from cryopreserved tissue chunk, as well as recovery of frozen isolated cells. We demonstrate cytoskeletal, ultrastructural, transcriptomic, metabolic, electrophysiological recovery of these cells both over short-term (1 week) and long-term (6–12 months) storage. In addition, these cells responded promptly to external stimuli, including adrenergic stimulation and hypoxic stress, suggesting functional integrity. Our study illustrates an optimized cryopreservation protocol for the functional recovery of cardiomyocytes from myocardial tissue, broadening their applications in basic and translational research, and opens up the possibility for cell banking.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105583"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146009177","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}
CryobiologyPub Date : 2026-03-01Epub Date: 2026-02-18DOI: 10.1016/j.cryobiol.2026.105597
Feiyu Peng , Lixiang Liu , Yifeng Yang , Xulin Zhang , Yushi Zhang , Bingfeng Fan , Xiaoyu Chu , Jie Cao , Ying Zhang , Weigang Zhao , Yu Tang , Baozeng Xu
{"title":"Glycine and betaine improve the quality of cryopreserved sika deer sperm","authors":"Feiyu Peng , Lixiang Liu , Yifeng Yang , Xulin Zhang , Yushi Zhang , Bingfeng Fan , Xiaoyu Chu , Jie Cao , Ying Zhang , Weigang Zhao , Yu Tang , Baozeng Xu","doi":"10.1016/j.cryobiol.2026.105597","DOIUrl":"10.1016/j.cryobiol.2026.105597","url":null,"abstract":"<div><div>Sperm cryopreservation imposes osmotic stress due to the addition and removal of high-concentration cryoprotectants. Whether organic osmolytes can alleviate this stress to improve cryopreservation efficiency remains unclear. Here, we evaluated the effects of glycine and betaine, supplemented as organic osmolytes throughout the cryopreservation process, on the quality and developmental potential of cryopreserved sika deer sperm. Pooled semen samples from five healthy adult stags were randomly allocated to four groups: a control (base extender) and three treatment groups supplemented with glycine and betaine at 6 mM + 200 μM, 10 mM + 500 μM, and 14 mM + 800 μM, respectively. The addition of 10 mM glycine and 500 μM betaine significantly improved post-thaw sperm quality, including enhanced total motility (TM), progressive motility (PM), average path velocity (VAP), curvilinear velocity (VCL), plasma membrane and acrosome integrity, and fertilization capacity, while reducing reactive oxygen species (ROS) levels compared with the control and other treatments. These findings suggest that glycine and betaine at 10 mM and 500 μM, respectively, may mitigate cryo-injury and enhance cryosurvival in sika deer sperm, offering a promising strategy for improving semen cryopreservation and artificial insemination protocols.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"122 ","pages":"Article 105597"},"PeriodicalIF":2.1,"publicationDate":"2026-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146225908","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}