Cryobiology最新文献

{"title":"Effects of co-supplementation of trehalose with fetuin and taxifolin in freezing extender on quality parameters of bull spermatozoa following freeze-thaw process","authors":"Mustafa Numan Bucak ,&nbsp;Morteza Taravat ,&nbsp;Oya Korkmaz ,&nbsp;Ali Soliemanzadeh ,&nbsp;Mojtaba Karimpour ,&nbsp;Mohammad Saber Maroufi ,&nbsp;Neda Abedpour ,&nbsp;Sadık Küçükgünay ,&nbsp;Tohid Rezaei Topraggaleh","doi":"10.1016/j.cryobiol.2025.105330","DOIUrl":"10.1016/j.cryobiol.2025.105330","url":null,"abstract":"<div><div>Glycerol is commonly used as a cryoprotectant in bovine sperm cryopreservation; however, its cytotoxic effects can negatively impact sperm quality. This study aimed to reduce glycerol concentration in semen extenders by supplementing with natural antioxidants (taxifolin and fetuin) and the disaccharide trehalose. Twelve ejaculates were collected from three bulls and initially evaluated, then diluted in a tris-egg yolk (TEY) extender containing either 7 % or 4 % glycerol. The control group received 7 % glycerol (TEY), while experimental groups included G7F (TEY + fetuin), G7T (TEY + taxifolin), G7M (TEY + fetuin + taxifolin), and 4 % glycerol groups supplemented with 50 mM trehalose: G4 (TEY + trehalose), G4F (TEY + fetuin), G4T (TEY + taxifolin), and G4M (TEY + fetuin + taxifolin). Post-thaw evaluations included assessments of sperm motility, plasma membrane integrity, acrosome integrity, mitochondrial membrane potential (MMP), DNA fragmentation, and oxidative stress markers. The G4F and G4T groups exhibited significantly higher total and progressive motility, viability, plasma membrane integrity, and MMP compared to other groups (p &lt; 0.05). Furthermore, these groups showed significantly lower malondialdehyde (MDA) levels, indicating reduced lipid peroxidation and oxidative stress (p &lt; 0.05). In conclusion, co-supplementation of 50 mM trehalose with 50 μM taxifolin and 5 mg/mL fetuin in a cryopreservation extender containing 4 % glycerol may enhance the post-thaw quality of bull spermatozoa.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"121 ","pages":"Article 105330"},"PeriodicalIF":2.1,"publicationDate":"2025-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145225103","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":"Research on the role of endoplasmic reticulum-mitochondria interactions in maintaining calcium homeostasis in skeletal muscle fibers of hibernating Daurian ground squirrels","authors":"Jie Zhang ,&nbsp;Fangyang Pan ,&nbsp;Yue He ,&nbsp;Jing Zhao ,&nbsp;Xiaozhuo Bai ,&nbsp;Yanhong Wei ,&nbsp;Yunfang Gao","doi":"10.1016/j.cryobiol.2025.105337","DOIUrl":"10.1016/j.cryobiol.2025.105337","url":null,"abstract":"<div><div>The ability to maintain cytoplasmic Ca²⁺ homeostasis during the torpor-arousal cycle is a crucial strategy that enables hibernators to resist disuse-induced muscle atrophy during hibernation. However, whether ER-mitochondria interactions contribute to the regulation of cytoplasmic Ca²⁺ homeostasis in skeletal muscle fibers during hibernation remain unclear. Here, we systematically explored the expression of tethering proteins, Ca²⁺ axis-related proteins, and protein complexes involved in Ca²⁺ transport within the ER-mitochondria contact sites (ERMCS) of soleus (SOL) and extensor digitorum longus (EDL) muscles in Daurian ground squirrels (<em>Spermophilus dauricus</em>) during hibernation. Results demonstrated a significant increase in the expression of the tethering protein MFN2 and Ca²⁺ axis-related proteins GRP75 and VDAC1, as well as enhanced interactions within the Ca²⁺ axis complex in the SOL muscle in the inter-bout arousal (IBA) group, as compared to the late torpor (LT) group. In addition, these same indicators exhibited no significant alternation in the EDL muscle during the torpor-arousal cycle. These findings suggest that ER-mitochondria interactions may participate in maintaining Ca²⁺ homeostasis in the skeletal muscles during the torpor-arousal cycle, especially in slow-twitch muscle.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"121 ","pages":"Article 105337"},"PeriodicalIF":2.1,"publicationDate":"2025-10-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145225095","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":"Corrigendum to \"Dormant bud cryopreservation of Rubus idaeus L.\" [Cryobiology 119 (2025) 105240].","authors":"Olena Bobrova, Jiri Zamecnik, Milos Faltus, Alois Bilavcik","doi":"10.1016/j.cryobiol.2025.105331","DOIUrl":"https://doi.org/10.1016/j.cryobiol.2025.105331","url":null,"abstract":"","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105331"},"PeriodicalIF":2.1,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145198699","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":"Papaverine enhances sperm quality in asthenospermia by reducing oxidative stress and inflammation while regulating the expression of apoptosis-related genes during cryopreservation","authors":"Zahra Azizi ,&nbsp;Malek Soleimani Mehranjani ,&nbsp;Mohammad Ali Shariatzadeh ,&nbsp;Nazila Najdi ,&nbsp;Atena Sadat Azimi","doi":"10.1016/j.cryobiol.2025.105329","DOIUrl":"10.1016/j.cryobiol.2025.105329","url":null,"abstract":"<div><div>Sperm motility is crucial for male fertility, allowing sperm to reach and fertilize the oocyte. Asthenozoospermia, marked by reduced motility, leads to infertility via oxidative stress, DNA damage, and mitochondrial dysfunction. Freeze-thawing, used in assisted reproductive technologies (ART), damages sperm. This study examined papaverine's antioxidant effects on improving asthenozoospermic sperm quality during freeze-thawing. Thirty semen samples from asthenozoospermic patients were divided into three groups: Control (fresh), Freeze (cryoprotectant, 30-min incubation), and Freeze + Papaverine (cryoprotectant + 100 μM papaverine, 30-min incubation). Sperm parameters, DNA fragmentation, TNF-α, IL-10, reactive oxygen species (ROS), malondialdehyde (MDA), total antioxidant capacity (TAC), antioxidant enzyme activities (SOD, CAT, GSH), plasma membrane integrity, mitochondrial membrane potential (MMP), and Bcl-2, Bax, and Caspase-3 expression at gene and protein levels were analyzed in all groups. In the Freeze group, sperm parameters, antioxidant enzyme levels, plasma membrane integrity, MMP, acrosomal integrity, levels of IL-10, and <em>Bcl-2</em> expression significantly decreased (p &lt; 0.05–0.001) compared to controls. In contrast, ROS, MDA, TNF-α, DNA fragmentation, and <em>Bax</em> and <em>Caspase-3</em> expression significantly increased (p &lt; 0.05–0.001). However, in the Freeze + Papaverine group, all these factors, except acrosomal integrity, showed significant improvement compared to the Freeze group. This study suggests that treating asthenozoospermic sperm with papaverine before freezing improves post-thaw sperm quality by enhancing motility, viability, plasma membrane integrity, and MMP, while reducing oxidative stress, inflammation, and apoptosis—potentially contributing to improved outcomes in ART.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"121 ","pages":"Article 105329"},"PeriodicalIF":2.1,"publicationDate":"2025-09-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145198690","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":"Editorial: Plant cryobiotechnology: multidisciplinary and modern research to understand the fundamental basis that allows successful cryopreservation and recovery.","authors":"Daniel Ballesteros","doi":"10.1016/j.cryobiol.2025.105328","DOIUrl":"https://doi.org/10.1016/j.cryobiol.2025.105328","url":null,"abstract":"<p><p>This Editorial introduces and contextualizes the papers of the Virtual Special Issue: Plant cryobiotechnology: multidisciplinary and modern research to understand the fundamental basis that allows successful cryopreservation and recovery.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105328"},"PeriodicalIF":2.1,"publicationDate":"2025-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145184948","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":"Exogenous abscisic acid enhances freeze-thaw stress tolerance in Antarctic moss Sanionia uncinata through coordinated antioxidant defense and osmoprotectant accumulation","authors":"Suyeon Seo ,&nbsp;Yong-Yoon Ahn ,&nbsp;Kitae Kim ,&nbsp;Jun Hyuck Lee ,&nbsp;Kyungwon Min ,&nbsp;Hyoungseok Lee","doi":"10.1016/j.cryobiol.2025.105325","DOIUrl":"10.1016/j.cryobiol.2025.105325","url":null,"abstract":"<div><div>Abscisic acid (ABA) is known to improve plant freeze-thaw stress tolerance (FTST), but its role in Antarctic mosses remains largely unexplored. Accordingly, this study investigated the effects of exogenous ABA application on FTST of gametophore <em>Sanionia uncinata</em>, one of the most widespread moss species in maritime Antarctica. Samples were treated with 10 μM ABA for 72 h under controlled culture conditions, then subjected to freezing stress at −13 and −16 °C. Various physiological parameters were measured to assess ABA-induced FTST, including ion-leakage, photosynthetic efficiency (Fv/Fm), malondialdehyde (MDA) content, compatible solutes/osmolytes (proline and total sugars), and antioxidant enzyme activities, e.g., superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX). Gametophores treated with ABA efficiently assimilated ABA, evident by ∼20-fold higher than controls. ABA application did not impede gametophore growth but slightly increased dry weight/fresh weight ratio (5 % increase). ABA significantly improved freezing tolerance, reducing freeze-induced membrane injury by ∼20 % at both stress temperatures, alleviating oxidative stress with 25–34 % lower MDA accumulation, and enhancing PSII maximum quantum yield (Fv/Fm) by 6–7 % following freeze-thaw stress. Improved FTST was paralleled by enhanced activities of antioxidant enzymes, CAT and APX increasing by ∼11 % and 56 %, respectively, while SOD remained unchanged. ABA treatment also promoted osmoprotectant accumulation, with proline content increasing by 33.5 % and total soluble sugars nearly doubling (99 % increase). These findings provide the first quantitative evidence that exogenous ABA markedly improves FTST in an Antarctic moss through coordinated antioxidant defense and osmoprotectant accumulation, highlighting ABA as a critical regulator of stress resilience in <em>S. uncinata</em>.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"121 ","pages":"Article 105325"},"PeriodicalIF":2.1,"publicationDate":"2025-09-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145148072","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":"Interrupted slow cooling of mouse cardiac endothelial cell monolayers","authors":"Leah A. Marquez-Curtis ,&nbsp;Janet A. W. Elliott","doi":"10.1016/j.cryobiol.2025.105319","DOIUrl":"10.1016/j.cryobiol.2025.105319","url":null,"abstract":"<div><div>The increasing evidence of the role of the endothelium in cardiac physiology and the importance of a functional endothelium for vascular grafts necessitate in vitro models for research. Cryopreservation of cardiac endothelial cell monolayers will enable their on-demand availability. Here, we used interrupted slow cooling (graded freezing) to characterize the cryobiological response of a mouse cardiac endothelial cell (MCEC) line in a monolayer format and describe a procedure for its successful cryopreservation. MCEC monolayers on fibronectin-coated coverslips made of Rinzl, whose coefficient of thermal expansion is matched to ice, were equilibrated at −5 °C, ice-nucleated, cooled at 1 °C/min, and at various temperatures during cooling, samples were either directly-thawed or plunged into liquid nitrogen and then thawed. In the absence of cryoprotectants, direct-thaw samples revealed cryoinjury from solute effects, while plunge-thaw samples revealed damage from intracellular ice formation. MCEC monolayers were then loaded with 5 % dimethyl sulfoxide (Me₂SO), 6 % hydroxyethyl starch (HES), and 2 % chondroitin sulfate (CS) prior to interrupted slow cooling. The maximum immediate post-thaw relative viability (93.7 ± 5.5 %) and absolute viability (95.5 ± 21.1 %) were attained after plunge from −40 °C. Both fresh and cryopreserved MCEC monolayers exhibited metabolic activity, angiogenic potential through tube formation on Matrigel, intracellular calcium signaling, and expression of the tight junction protein ZO-1. Viability and functional assessments after post-thaw overnight culture did not show delayed-onset cryoinjury. Thus, the combination of Me₂SO, HES, CS, and Rinzl substrate resulted in viable and functional MCEC monolayers post thaw. This cryopreservation protocol will allow accessibility of cardiac endothelial monolayers for use in studying cardiac physiology, cardiovascular disease modeling, and drug-induced cardiotoxicity testing.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"121 ","pages":"Article 105319"},"PeriodicalIF":2.1,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145119199","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":"Screening for cryoprotective agent toxicity and toxicity reduction in mixtures at subambient temperatures","authors":"Nima Ahmadkhani ,&nbsp;Cameron Sugden ,&nbsp;Adam T. Mayo,&nbsp;Adam Z. Higgins","doi":"10.1016/j.cryobiol.2025.105315","DOIUrl":"10.1016/j.cryobiol.2025.105315","url":null,"abstract":"<div><div>Organ transplantation faces major challenges in preserving and transporting organs due to the limitations of existing cold storage methods. Cryopreservation offers a promising alternative, but it remains a challenge to avoid toxicity from the high concentrations of cryoprotective agents (CPAs) required to prevent ice formation. In this study, we used bovine pulmonary artery endothelial cells as a model system to systematically investigate CPA toxicity. To this end, we upgraded our high-throughput CPA toxicity screening platform by adding subambient temperature control, enabling assessment of CPA toxicity at 4 °C, a temperature commonly used for CPA equilibration in tissue and organ cryopreservation. Overall, we screened 22 individual CPAs and a wide range of binary mixtures at concentrations up to 12 mol/kg, allowing us to identify CPA combinations that reduce toxicity. Our findings revealed that at 4 °C, CPA toxicity was significantly reduced compared to room temperature. Several CPA combinations resulted in significantly lower toxicity than their constituent CPAs at the same concentration, including 12 CPA mixtures at 6 mol/kg and 8 CPA mixtures at 12 mol/kg. Toxicity neutralization was also observed in 9 cases, especially in combinations involving formamide, acetamide, dimethyl sulfoxide, and glycerol. For example, exposure to 6 mol/kg formamide alone resulted in 20 % viability, but the addition of 6 mol/kg glycerol to create a mixture with a total concentration of 12 mol/kg eliminated this toxicity, resulting in a viability of 97 %. These findings support the rationale for using multi-CPA cocktails and underscore the potential of rational mixture design to reduce toxicity.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"121 ","pages":"Article 105315"},"PeriodicalIF":2.1,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145119200","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":"Retrospective analysis of sperm metrics in fresh ejaculates of collared peccaries presenting contrasting semen freezability","authors":"Samara Sandy Jerônimo Moreira ,&nbsp;Romário Parente dos Santos ,&nbsp;Luana Grasiele Pereira Bezerra ,&nbsp;Ana Glória Pereira ,&nbsp;Náyra Rachel Nascimento Luz ,&nbsp;Andreza Vieira Brasil ,&nbsp;Thales Pinheiro Cavalcanti ,&nbsp;Moacir Franco de Oliveira ,&nbsp;Simone Maria Massami Kitamura Martins ,&nbsp;André Furugen Cesar de Andrade ,&nbsp;Alexandre Rodrigues Silva","doi":"10.1016/j.cryobiol.2025.105324","DOIUrl":"10.1016/j.cryobiol.2025.105324","url":null,"abstract":"<div><div>Based on a retrospective study of sperm parameters in raw ejaculates of collared peccaries classified as good and poor freezers, this study aimed to identify whether these animals present different sperm characteristics that allow predicting their contrasting freezability. For this purpose, ejaculates were obtained from twenty-six animals, in which an aliquot of fresh semen was immediately evaluated, while another aliquot was diluted in Tris-yolk-glycerol diluent and cryopreserved. Fresh and frozen/thawed samples were evaluated for sperm motility kinetic parameters, membrane integrity, mitochondrial activity, membrane functionality, sperm morphology and sperm binding capacity. Based on post-thawing sperm metrics, the animals were classified as good and poor freezers. Following this classification, a retrospective study of fresh ejaculates was performed, in which a reclassification of the samples was performed according to the groups obtained after thawing. For the thawed groups, analysis of variance was performed using SAS software and the Tukey mean test to detect the significant differences between the groups. For fresh samples, the Mann Whitney test was used to verify the difference between the groups regarding the kinetic motility parameters (MT, fast, medium, slow and static cells), mitochondrial activity, pH and sperm concentration. For the other fresh parameters, the Unpaired t-test was used, using the GraphPad Prism® software version 9.3 for Windows (GraphPad Software Inc., San Diego, CA, USA). The highest (P &lt; 0.05) mean values of post-thawing sperm motility were verified for good freezers (42.8 ± 2.8 %), to the detriment of bad freezers (23.8 ± 2.7 %). Good freezers also showed the highest (P &lt; 0.05) mean values of progressive sperm motility (25.8 ± 2.5 %), lateral head amplitude (6.1 ± 0.1 μm) and proportion of fast sperm (31.1 ± 2.6 %), while poor freezers showed the highest proportion (P &lt; 0.05) of static sperm (72.8 ± 3.4 %). Regarding raw semen, all ejaculates had a milky appearance and whitish coloration. There was no statistical difference between the groups regarding the kinetic motility patterns (P &gt; 0.05). Poor freezers provided the lowest percentage (P &lt; 0.05) of sperm with intact membranes (78.8 ± 1.7 %) and mitochondrial activity (79.3 ± 1.8 %), compared to good freezers (87.0 ± 1.9 %; 87.3 ± 1.9 %), with no differences regarding the other parameters. In conclusion, sperm plasma membrane integrity and mitochondrial activity in raw semen appear to be useful for predicting the freezability of semen from collared peccaries classified as good and poor freezers.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"121 ","pages":"Article 105324"},"PeriodicalIF":2.1,"publicationDate":"2025-09-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145106496","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":"High-throughput evaluation of cryoprotective agents for mixture effects that reduce toxicity","authors":"Nima Ahmadkhani ,&nbsp;Cameron Sugden ,&nbsp;James D. Benson ,&nbsp;Ali Eroglu ,&nbsp;Adam Z. Higgins","doi":"10.1016/j.cryobiol.2025.105316","DOIUrl":"10.1016/j.cryobiol.2025.105316","url":null,"abstract":"<div><div>Vitrification is a promising approach for cryopreserving complex biological structures such as organs. However, to prevent ice formation, high concentrations of cell-permeable cryoprotective agents (CPAs) are required, which can be highly toxic. The current reliance on a small number of CPAs limits optimization of low-toxicity compositions for vitrification. To address this, there is growing interest in uncovering novel chemicals with analogous protective qualities. This may not only enhance vitrification efficacy but also mitigate toxic effects. In the current study, we employed a high throughput method to assess the toxicity of 21 compounds at room temperature, both individually and in binary combinations, using bovine pulmonary artery endothelial cells (BPAEC) as a model system. Our analysis revealed that toxicity increases with both exposure duration and concentration, and that several CPA combinations result in reduced overall toxicity. Notably, among all tested mixtures, four binary combinations—formamide/glycerol, dimethyl sulfoxide/1,3-propanediol, 1,2-propanediol/diethylene glycol, and 1,3-propanediol/diethylene glycol—produced a statistically significant decrease in toxicity, resulting in significantly higher viability for the 6 mol/kg mixture than both corresponding 6 mol/kg single CPA solutions. The high-throughput approach presented here will aid in building a comprehensive CPA toxicity database, which will improve our understanding of toxicity mechanisms and support the development of predictive models for identifying novel CPA mixtures with low toxicity.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"121 ","pages":"Article 105316"},"PeriodicalIF":2.1,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145079834","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}