Cryobiology最新文献

英文中文

Extracellular vesicles as emerging bioactive modulators in sperm cryopreservation: Mechanisms and therapeutic potentials 细胞外囊泡作为精子冷冻保存中新兴的生物活性调节剂：机制和治疗潜力

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-13 DOI: 10.1016/j.cryobiol.2025.105568

Merve Deniz Tanrıkulu , Mesut Çevik , Mustafa Numan Bucak , Alper Koçyiğit

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.

精子冷冻保存是辅助生殖和遗传保护的基础，但冷冻解冻过程会对精子造成严重的结构和功能损伤。该领域的最新进展将细胞外囊泡（EVs）定位为潜在的治疗工具，主要是因为它们具有运输功能性生物分子的能力，有助于抵消低温保存引起的细胞损伤。然后，我们研究了精子冷冻保存过程中遇到的细胞和分子损伤类型，并综合了ev在这种情况下的保护作用的现有证据。特别关注ev可能保持精子完整性的机制，包括膜稳定、抗氧化传递和信号通路的调节。尽管有令人鼓舞的发现，但挑战依然存在，包括电动汽车隔离方法的标准化、最佳给药和给药策略。我们还讨论了作用机制研究和多组学方法的必要性，以更好地了解ev -精子相互作用。最后，展望了未来的发展方向，包括电动汽车工程和临床翻译。总的来说，ev在低温生物学中成为有前途的调节剂，具有改善精子保存结果和生育恢复策略的潜力。本文综述了ev的深入研究，包括其形成过程、分离ev的各种策略以及用于评估ev性质的技术。它还强调了它们在男性生殖系统中的来源和功能意义。

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引用次数: 0

Coaxial temperature controlled cryoprinting: A biomimetic technology inspired by the freezing survival mechanisms of the frog Rana sylvatica 同轴温度控制冷印：一种仿生技术，灵感来自蛙蛙的冷冻生存机制

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-13 DOI: 10.1016/j.cryobiol.2025.105569

Leo Lou , Linnea Warburton , Cristina Bilbao-Sainz , Nicholas Jean , Boris Rubinsky

Inspired by the freeze-survival mechanism of the frog Rana sylvatica, we developed a co-axial, self-crosslinking approach to be used when fabricating frozen scaffolds. Soft biomaterial can be fabricated into scaffolds using methods such as Temperature Controlled Cryoprinting (TCC), which uses a freezing plate to freeze the biomaterial as it is extruded. However, it remains a challenge to uniformly crosslink the scaffolds during thawing without them losing their shape. In this paper, we use coaxial printing to generate locally variable phase-transition compositions, mimicking the frog's glucose distribution during freezing, and electrical Joule heating to simulate metabolic heat production. Unlike previous methods, in our novel approach the crosslinker is located already within the frozen scaffold, and there is no need to submerge the scaffold in a bath. Both mathematical modeling and experimental validation confirmed the concept. A finite thawing-diffusion-crosslinking model was developed to analyze parameters such as temperature, thawing interface, concentration, and crosslinking interface. Experimental evaluations, including mechanical testing, heating test, swelling performance and electron microscope, demonstrated that coaxial self-crosslinking enables controlled thawing and crosslinking of frozen scaffolds. Results demonstrated that coaxial TCC self-crosslinking scaffolds are feasible, allowing for a more controlled thawing and crosslinking process in cryoprinted scaffolds.

受到蛙蛙（Rana sylvatica）冷冻存活机制的启发，我们开发了一种同轴、自交联的方法，用于制造冷冻支架。柔软的生物材料可以通过温度控制冷冻打印（TCC）等方法制成支架，TCC使用冷冻板在挤压生物材料时将其冷冻。然而，如何在解冻过程中使支架均匀交联而不失去其形状仍然是一个挑战。在本文中，我们使用同轴打印来生成局部可变的相变成分，模拟青蛙在冷冻过程中的葡萄糖分布，并使用电焦耳加热来模拟代谢热的产生。与以前的方法不同，在我们的新方法中，交联剂已经位于冷冻支架内，并且不需要将支架浸入浴缸中。数学模型和实验验证都证实了这一概念。建立了有限融化-扩散-交联模型，分析了温度、融化界面、浓度和交联界面等参数。包括力学测试、加热测试、膨胀性能和电子显微镜在内的实验评估表明，同轴自交联可以实现冷冻支架的可控解冻和交联。结果表明，同轴TCC自交联支架是可行的，允许在冻印支架中更可控的解冻和交联过程。

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引用次数: 0

Modulation of inflammation of lipid metabolism by sesamin, forskolin, α-linolenic acid, and their combination in a zebrafish model of cryo-injury and high cholesterol diet-induced myocardial infarction, followed by atherosclerosis 芝麻素、福斯克林、α-亚麻酸及其组合在低温损伤和高胆固醇饮食诱导的心肌梗死后动脉粥样硬化斑马鱼模型中的脂质代谢炎症调节作用

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-12 DOI: 10.1016/j.cryobiol.2025.105565

Abu Safana Biswas, Kamsagara Linganna Krishna, Ganavi Bethanagere Ramesha, Pooja Gandharvachari Achar

Cryo-injury has been a standard model used to simulate myocardial infarction (MI), which may induce reproducible myocardial necrosis under controlled conditions. In a Zebrafish (ZF) model of cryoinjury and a high-cholesterol diet (HCD), the study evaluates the preventive effects of sesamin, forskolin, and α-linolenic acid, either alone or in combination, against myocardial infarction with atherosclerosis. Various physiological, biochemical, and histological endpoints were used to evaluate the effectiveness of the treatment. A notable recovery of cardiac activity (as measured by ECG) and body weight regulation was observed when comparing the treated groups with the cryo-injured control. Decreased TNF-α and iNOS levels demonstrated effective molecular control of cryoinjury-induced inflammation. An improved lipid balance was also observed through lipid profiling of cardiac tissue and serum, which revealed significant increases in HDL levels and decreases in triglycerides, total cholesterol, LDL, and VLDL. The treatment with a combination of test compounds especially shows good efficacy towards myocardial infarction and the myocardial structure of ZF, followed by atherosclerosis. In our study, the combination effect of sesamin, forskolin, and α-linolenic acid has significant potential for regulating lipid levels, inflammatory markers, and myocardial structure integrity. It can be concluded that the combination treatment of the sesamin, forskolin, and α-linolenic acid may protect the cardiovascular system, and this combination can be used for further preclinical study with a rodent model for cardiovascular disease.

低温损伤已成为模拟心肌梗死（MI）的标准模型，在可控条件下可诱发可重复性心肌坏死。在冷冻损伤和高胆固醇饮食（HCD）的斑马鱼（ZF）模型中，研究评估了芝麻素、福斯克林和α-亚麻酸单独或联合使用对心肌梗死合并动脉粥样硬化的预防作用。使用各种生理、生化和组织学终点来评估治疗的有效性。与低温损伤对照组相比，治疗组心脏活动（ECG测量）和体重调节明显恢复。TNF-α和iNOS水平的降低显示了低温损伤诱导炎症的有效分子控制。通过心脏组织和血清的脂质分析也观察到脂质平衡的改善，这表明HDL水平显著增加，甘油三酯、总胆固醇、LDL和VLDL水平降低。试验性化合物联合治疗尤其对心肌梗死和ZF心肌结构有较好的疗效，其次是动脉粥样硬化。在我们的研究中，芝麻素、福斯克林和α-亚麻酸的联合作用在调节血脂水平、炎症标志物和心肌结构完整性方面具有显著的潜力。由此可见，芝麻素、福斯克林和α-亚麻酸联合用药对心血管系统有一定的保护作用，该联合用药可用于啮齿动物心血管疾病模型的临床前研究。

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引用次数: 0

Numerical thermal analysis and optimization of vitrification cooling strategies for porcine heart cryopreservation 猪心脏冷冻玻璃化冷却策略的数值热分析与优化

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-11 DOI: 10.1016/j.cryobiol.2025.105570

Wangjie Sun , Xueqiang Dong , Xian Wang , Pengsong Yuan , Maoqiong Gong

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⁻¹, and a convective heat transfer coefficient of 250 W∙m⁻²∙°C⁻¹, 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⁻¹, surpassing the critical cooling rate of VS55 + 0.6 M sucrose (<1.0 °C·min⁻¹), 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.

基于玻璃化的低温保存为器官的长期保存提供了一种很有前途的方法；然而，在大体积器官中实现均匀冷却仍然是一个重大挑战，因为严重的热梯度会导致这些器官内的开裂和结冰。利用基于MRI数据的猪心脏高保真三维有限元模型，我们系统地研究了初始冷却温度、玻璃相冷却速率、对流换热系数、冷冻保护剂（CPA）导热系数和容器几何形状等关键参数对玻璃化性能的影响。仿真结果表明，优化方案的初始冷却温度为−10°C，玻璃相冷却速率为1.0°C·min−1，对流换热系数为250 W∙m−2∙°C−1，成功地平衡了冷却效率和热均匀性。在此条件下，心脏组织和CPA的平均中央冷却速率均超过1.4°C·min - 1，超过了VS55 + 0.6 M蔗糖的临界冷却速率（<1.0°C·min - 1），同时将内部温差峰值限制在25.5°C以下，以最小化热应力。此外，我们发现提高CPA导热系数，降低其比热容，并使用圆柱形容器显著改善了温度均匀性，缩短了平衡时间。这些发现为优化玻璃化策略提供了重要的定量见解，并为设计更安全、更有效的大器官临床冷冻保存方案奠定了坚实的理论基础。

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引用次数: 0

Antioxidant astaxanthin enhances cryosurvival and post-thaw functional parameters of rabbit spermatozoa 抗氧化剂虾青素可提高兔精子的冷冻存活和解冻后功能参数

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-11 DOI: 10.1016/j.cryobiol.2025.105564

Paula Luque , Lenka Kuželová , Jakub Vozaf , Andrej Baláži , Peter Chrenek

This study evaluated the effect of astaxanthin (AX), a potent antioxidant carotenoid, on the post-thaw quality of cryopreserved rabbit sperm. Semen samples were frozen with AX supplementation at concentrations of 0 (CONTROL), 0.5, 1, or 2 μM. Post-thaw sperm quality was assessed by computer-assisted sperm analysis (CASA) for motility, and by flow cytometry to evaluate viability, early apoptosis, mitochondrial activity, reactive oxygen species (ROS) levels, and acrosomal membrane integrity. Supplementation with 0.5 and 1 μM AX significantly improved total motility, viability, and mitochondrial activity compared to the control group (p < 0.05). These concentrations also led to significantly reduced levels of apoptotic cells and ROS. Acrosomal damage was not significantly affected by AX supplementation. These findings demonstrate that low-dose AX addition during cryopreservation attenuates oxidative and apoptotic damage in rabbit spermatozoa and enhances several key post-thaw quality parameters. Astaxanthin may thus represent a promising additive for improving cryosurvival in rabbit sperm used for assisted reproduction.

研究了虾青素（AX）对兔精子解冻后质量的影响。虾青素是一种有效的抗氧化类胡萝卜素。精液样品在0（对照）、0.5、1或2 μM浓度的AX中冷冻。通过计算机辅助精子分析（CASA）评估解冻后精子的活力，并通过流式细胞术评估活力、早期凋亡、线粒体活性、活性氧（ROS）水平和顶体膜完整性。与对照组相比，添加0.5和1 μM AX显著提高了总运动性、活力和线粒体活性（p < 0.05）。这些浓度也导致凋亡细胞和ROS水平显著降低。添加AX对顶体损伤无显著影响。这些结果表明，在冷冻保存期间添加低剂量的AX可减轻兔精子的氧化和凋亡损伤，并提高解冻后的几个关键质量参数。虾青素可能是一种很有前途的添加剂，可以提高兔精子的冷冻存活率，用于辅助生殖。

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引用次数: 0

Cryopreservation of Drosophila melanogaster embryonic nuclei in a Dimethyl sulfoxide (ME2SO)-free solution 黑腹果蝇胚胎核在无二甲亚砜（ME2SO）溶液中的低温保存

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-10 DOI: 10.1016/j.cryobiol.2025.105561

Troy Louwagie , Lindsey G. O’Brien , Daryl M. Gohl , Allison Hubel

Drosophila melanogaster is a well-known organism for biological research. With more than 150,000 live stocks maintained, there is an urgent need for cryopreservation of Drosophila stocks to limit maintenance costs and genetic drift. Two methods have been described for freezing Drosophila stocks: vitrification of embryos and of primordial germ cells. These methods show success in cryopreserving stocks, but the robustness and the applicability of these approaches in preserving a wide range of mutant stocks has not been established. Here, we describe a method for cryopreserving isolated embryonic nuclei using a slow-cooling method. Using a differential evolution algorithm, we identified five different formulations that show

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80% recovery and used Raman spectroscopy to image nuclei at low temperatures. Finally, we demonstrate that cryopreserved nuclei retain biological function through an induced hsp70 transcriptional response. This work represents steps towards cryopreserving nuclei with the ultimate goal of regenerating stocks by embryonic nuclear transplantation.

黑腹果蝇是生物学研究中著名的生物。由于维持了超过150,000个活的果蝇种群，因此迫切需要对果蝇种群进行低温保存，以限制维持成本和遗传漂变。已经描述了两种冷冻果蝇种群的方法：胚胎玻璃化和原始生殖细胞玻璃化。这些方法在冷冻保存中取得了成功，但这些方法在保存大范围突变体砧木方面的稳健性和适用性尚未建立。在这里，我们描述了一种使用缓慢冷却方法冷冻保存离体胚胎核的方法。使用差分进化算法，我们确定了五种不同的配方，显示80%的回收率，并使用拉曼光谱在低温下对原子核进行成像。最后，我们证明低温保存的细胞核通过诱导hsp70转录反应保留了生物学功能。这项工作代表了细胞核低温保存的最终目标是通过胚胎核移植再生砧木的步骤。

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引用次数: 0

Development of B cell medicine cryopreservation process B细胞药物冷冻保存工艺的研究进展

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-01 DOI: 10.1016/j.cryobiol.2025.105366

Mark Lalli, Jordan Smelker, Rashmi Kshirsagar

引用次数: 0

Improving non-contact cooling rate with liquid nitrogen by hierarchical surface structures 利用分层表面结构提高液氮非接触冷却速率

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-01 DOI: 10.1016/j.cryobiol.2025.105371

Meng Shi

引用次数: 0

Permeable and non-permeable cryoprotectant-free cryopreservation of larger volumes human semen 大容量人精液的透性和不透性无冷冻保护剂冷冻保存

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-01 DOI: 10.1016/j.cryobiol.2025.105359

Karekin D. Esmeryan , Todor A. Chaushev

引用次数: 0

Oocyte and embryo vitrification using automated equipment based on microfluidic chip 基于微流控芯片的自动化设备对卵母细胞和胚胎进行玻璃化冷冻

IF 2.1 3区生物学 Q2 BIOLOGY

Cryobiology

Pub Date : 2025-12-01 DOI: 10.1016/j.cryobiol.2025.105360

Jing Shen , Yukun Cao , Weijie Li , Xinli Zhou

引用次数: 0

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