Cryobiology最新文献

{"title":"Long-term hypothermic storage of oocytes of the European common frog Rana temporaria at various pressure regimes in gas mixtures based on oxygen, carbon monoxide, and nitrous oxide","authors":"Evgeniy L. Gagarinskiy,&nbsp;Viktor K. Uteshev,&nbsp;Eugeny E. Fesenko Jr.","doi":"10.1016/j.cryobiol.2024.104952","DOIUrl":"10.1016/j.cryobiol.2024.104952","url":null,"abstract":"<div><p>In recent years, the challenge of preserving amphibian biodiversity has increasingly been addressed through technologies for the short-term storage of unfertilized spawn at low positive temperatures. Previously the possibility of using a 6.5 atm gaseous mixture of carbon monoxide and oxygen for prolonged hypothermic preservation of unfertilized oocytes for more than 4 days was shown. This study aimed to investigate the viability of oocytes R. temporaria preserved under conditions of hypothermia at 2.5, 3 and 6.5 excess atm pressure in the various gas mixture compositions (CO, N₂O, O₂) and pure oxygen. The use of pressure up to 3 excess atmospheres was significantly beneficial compared to 6.5 atm at the 7 days storage period. The results indicate that oxygen pressure is a critical factor in maintaining oocyte viability. Admixing CO or N₂O to oxygen reduced variability in the results but did not significantly affect the measured indicators (fertilization, hatching) in the experimental groups. The composition CO + O₂ (0.5/3.5 ratio, 3 excess atm) reliably extended the shelf life of viable oocytes, indistinguishable from native controls by fertilization and hatching rates, to 4 days. After 7 days, oocytes exhibited fertilization and hatching rates that were 79 % and 48 % compared to native control. Reducing the pressure of the preserving gas mixture to 3 atm, as utilized in this study, simplifies the practical implementation of gas preservation technology for maintaining endangered amphibian species during breeding in laboratory conditions.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141916307","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":"Preparation of subcooled liquid argon and test of its cooling ability","authors":"Mingsheng Li ,&nbsp;Yifei Sun ,&nbsp;Xianguo Xu ,&nbsp;Shaozhi Zhang","doi":"10.1016/j.cryobiol.2024.104949","DOIUrl":"10.1016/j.cryobiol.2024.104949","url":null,"abstract":"<div><p>Subcooled liquid nitrogen and nitrogen slush are often considered for high-speed cooling, but their preparation and maintenance are not easy. To address this issue, a unique device was designed to prepare subcooled liquid argon (SLA) using liquid nitrogen (LN). The cooling process was mathematically modeled to predict the preparation time. If the interlayer space between LN and liquid argon is filled with nitrogen gas, liquid argon could be cooled to 3.5 K subcooling within 1 h. If the interlayer is filled with air, 2 h are required to achieve the same subcooled state. An additional 1000 mL of LN was required for the preparation of 600 mL of 3.5 K SLA. The cooling tests of 3 μL microdroplets in 3 mm–6 mm capillary quartz tubes were duplicated to evaluate the potential of SLA. It was found that the cooling rate of microdroplet in the 3.5 K subcooled SLA is very close to that in the 3 K subcooled LN, higher than that in the saturated LN. The convenience of preparation and maintenance of SLA can make it good choice of cryogen for cryopreservation of biomaterials.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141906173","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":"Factors affecting cryotolerance of mammalian oocytes","authors":"Lucia Olexiková ,&nbsp;Alexander Makarevich ,&nbsp;Linda Dujíčková ,&nbsp;Elena Kubovičová ,&nbsp;Peter Chrenek","doi":"10.1016/j.cryobiol.2024.104946","DOIUrl":"10.1016/j.cryobiol.2024.104946","url":null,"abstract":"<div><p>Cryopreservation of oocytes is an important tool for preserving genetic resources and for farm animals breeding. Processes taking place during vitrification affect oocytes and result in their reduced developmental capacity and lower fertilisation rates of cryopreserved oocytes. Further improvement in cryopreservation techniques is still required. Several authors already summarized the actual state and perspectives of oocyte cryopreservation as well as potential approaches to improve their development after thawing. The aim of this review is to specify factors affecting cryotolerance of mammalian oocytes, especially bovine <em>in vitro</em> matured oocytes, and to identify the areas, where more efforts were made to improve the success of oocyte cryopreservation. These factors include oocyte lipid content, membrane composition, mRNA protection, cytoskeleton stabilization and application of such potential stimulators of cell cryotolerance as antioxidants, growth factors or antifreeze proteins.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-08-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141787472","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of Vitis vinifera zygotic embryo cryopreservation and post-cryopreservation on the gene expression of DNA demethylases","authors":"Juan Luis García-Vázquez ,&nbsp;Mariana Quijada-Rivera ,&nbsp;Miguel Ángel Hernández-Oñate ,&nbsp;Martín Ernesto Tiznado-Hernández ,&nbsp;María Fernanda Lazo-Javalera ,&nbsp;Miguel Ángel Martínez-Téllez ,&nbsp;Karen Rosalinda Astorga-Cienfuegos ,&nbsp;Marisela Rivera-Domínguez","doi":"10.1016/j.cryobiol.2024.104947","DOIUrl":"10.1016/j.cryobiol.2024.104947","url":null,"abstract":"<div><p>Grapevine (<em>Vitis vinifera</em> L.) crops are continuously exposed to biotic and abiotic stresses, which can cause genetic and epigenetic alterations. To determine the possible effects of grapevine cryopreservation on the regulation of DNA demethylase genes, this work studied the expression of DNA demethylase genes in cryopreserved and post-cryopreserved grapevine tissues. <em>V. vinifera</em> DNA demethylases were characterized by <em>in silico</em> analysis, and gene expression quantification was conducted by RT‒qPCR. Three DNA demethylase sequences were found: VIT_13s0074g00450 (VvDMT), VIT_08s0007g03920 (VvROS1), and VIT_06s0061g01270 (VvDML3). Phylogenetic analysis revealed that the sequences from <em>V. vinifera</em> and <em>A. thaliana</em> had a common ancestry. In the promoters of responsive elements to transcription factors such as AP-2, Myb, bZIP, TBP, and GATA, the conserved domains RRM DME and Perm CXXC were detected. These responsive elements play roles in the response to abiotic stress and the regulation of cell growth. These data helped us characterize the <em>V. vinifera</em> DNA demethylase genes. Gene expression analysis indicated that plant vitrification solution 2 (PVS2) treatment does not alter the expression of DNA demethylase genes. The expression levels of VvDMT and VvROS1 increased in response to cryopreservation by vitrification. Furthermore, in post-cryopreservation, VvROS1 was highly induced, and VvDML3 was repressed in all the treatment groups. Gene expression differences between different treatments and tissues may play roles in controlling methylation patterns during gene regulation in tissues stressed by cryopreservation procedures and in the post-cryopreservation period during plant growth and development.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-08-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141859303","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":"Biodegradable capsules as a sustainable and accessible container for vitrification of gonadal tissue using the zebrafish animal model","authors":"Thaiza Rodrigues de Freitas ,&nbsp;Rômulo Batista Rodrigues ,&nbsp;Lis Santos Marques ,&nbsp;Renata Villar Dantas ,&nbsp;Karel Gelina Torres-Lozano ,&nbsp;Thales Souza França ,&nbsp;Larise Caroline Oliveira Lima ,&nbsp;Francielli Weber Santos ,&nbsp;Eduardo Thomé Nicoleti ,&nbsp;Tales Fabris Chaves ,&nbsp;Danilo Pedro Streit Jr","doi":"10.1016/j.cryobiol.2024.104944","DOIUrl":"10.1016/j.cryobiol.2024.104944","url":null,"abstract":"<div><p>Cryopreservation of fish gonadal tissue is an important technique for preserving genetic variability. However, this technique involves the use of cryotubes, plastic containers with low degradability that are expensive and difficult to obtain in certain parts of the world. Therefore, this study aimed to evaluate the efficiency of gelatin and hypromellose hard capsules as a sustainable and accessible alternative container to the cryotube for vitrification of zebrafish (<em>Danio rerio</em>) gonadal tissue. The gonadal tissues (testicular or ovarian) were vitrified in cryotubes, hard-gelatin, and hard-hypromellose capsules. Gelatin capsules exhibited comparable efficacy to cryotubes in preserving spermatogonia viability (33.03 ± 10.03 % and 37.96 ± 8.35 %, respectively), whereas hypromellose capsules showed decreased viability (18.38 ± 2.09 %). Immature oocyte viability remained unaffected by the capsule materials, with no difference compared to cryotubes at all oocyte stages (Primary Growth: p &lt; 0.0001; Cortical Alveolar: p &lt; 0.0001; Vitellogenic: p &lt; 0.0001). Mitochondrial activity and lipid peroxidation demonstrated no difference among cryotubes and capsules for both gonadal tissues. However, antioxidant activity was notably higher in gelatin capsules (Testes: 147.2 ± 32.32 μg; Ovary: 87.98 ± 10.91 μg) than in cryotubes (Testes: 81.04 ± 26.05 μg; Ovary: 54.35 ± 11.23 μg) and hypromellose capsules (Testes: 62.36 ± 17.10 μg; Ovary: 63.96 ± 7.51 μg), likely due to the inherent antioxidant properties of gelatin. The results obtained in this study demonstrate that the cryotube can be replaced by gelatin capsules for vitrification of both gonadal tissues of zebrafish, being a sustainable and accessible alternative as it is a low-cost and environmentally friendly container.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-08-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141733658","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":"Osmotic injury and cytotoxicity for hMSCs in contact with Me2SO: The effect of cell size distribution","authors":"Gabriele Traversari,&nbsp;Antonio Mario Locci,&nbsp;Alessandro Concas,&nbsp;Nicola Lai,&nbsp;Alberto Cincotti","doi":"10.1016/j.cryobiol.2024.104943","DOIUrl":"10.1016/j.cryobiol.2024.104943","url":null,"abstract":"<div><p>The paper discusses the impact of cell size on cytotoxicity and expansion lysis during the osmotic excursions resulting from the contact of hMSCs from UCB with Me2SO. It builds upon the mathematical model recently presented by the authors, which pertains to a population of cells with uniform size. The objective is to enhance the model's relevance by incorporating the more realistic scenario of cell size distribution, utilizing a Population Balance Equations approach. The study compares the capability of the multiple-sized model to the single-sized one to describe system behavior experimentally measured through cytofluorimetry and Coulter counter when, first, suspending hMSCs in hypertonic solutions of Me2SO (at varying osmolality, system temperature, and contact times), and then (at room temperature) pelleting by centrifugation before suspending the cells back to isotonic conditions. Simulations demonstrate that expansion lysis and cytotoxic effect are not affected by cell size for the specific system hMSCs/Me2SO, thus confirming what was found so far by the authors through a single-size model. On the other hand, simulations show that, when varying the adjustable parameters of the model that are expected to change from cell to cell lineages, expansion lysis is sensitive to cell size, while cytotoxicity is not, being mainly influenced by external CPA concentration and contact duration. More specifically, it is found that smaller cells suffer expansion lysis more than larger ones. The findings suggest that different cells from hMSCs may require a multiple-sized model to assess cell damage during osmotic excursions in cryopreservation.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-08-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0011224024000981/pdfft?md5=2c0fb1b626dd724daf90be749081b024&pid=1-s2.0-S0011224024000981-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141733660","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isolation and cryopreservation of Pseudopimelodus mangurus (Siluriformes) spermatogonial cells","authors":"Giselle Pessanha Pessoa ,&nbsp;Lucia Suárez López ,&nbsp;Jenyffer Mairely Rosero ,&nbsp;Silvio Carlos Alves dos Santos ,&nbsp;George Shigueki Yasui ,&nbsp;José Augusto Senhorini ,&nbsp;Paulo Sérgio Monzani","doi":"10.1016/j.cryobiol.2024.104941","DOIUrl":"10.1016/j.cryobiol.2024.104941","url":null,"abstract":"<div><p>Spermatogonia cryopreservation can be a strategy for future conservation actions. The neotropical Siluriformes <em>Pseudopimelodus mangurus</em> was already classified as vulnerable on the Red List of Threatened Species. <em>P. mangurus</em> spermatogonial cells were isolated, assessed, and cryopreserved. Fragments of the testis were enzymatically dissociated, purified using Percoll density gradient, and submitted to differential plating. Fractionated cells were evaluated by microscopy, <em>ddx4 (vasa)</em> relative expression, and alkaline phosphatase activity. Cryopreservation was conducted using ethylene glycol, glycerol, dimethyl sulfoxide (DMSO), dimethylacetamide (DMA), and propanediol at 1 M, 1.5 M, and 2 M. Cell viability was evaluated and cell concentration was determined. Cell fractions from 20 % and 30 % Percoll gradient bands showed the highest concentrations of spermatogonia. The fraction mix showed 54 % purity and 93 % viability. After differential plating, 60 % purity and 92 % viability were obtained. Spermatogonial cells showed high alkaline phosphatase activity compared to spermatocytes and spermatids. The relative spermatogonial <em>ddx4</em> expression from the Percoll density gradient was about twice as high as in samples from the testis and the differential plating. The increased <em>ddx4</em> expression indicated the enrichment of spermatogonial cells by density gradient step and dead cells expressing <em>ddx4</em> in differential plating, or <em>ddx4</em> decreasing expression during cell culture. For this reason, cells from the Percoll gradient were chosen for cryopreservation. Propanediol at 1 M demonstrated the best condition for spermatogonial cell cryopreservation, presenting 98 % viability, while dimethylacetamide at 2 M represented the least favorable condition, with approximately 47 % viability. These findings are essential for <em>P. mangurus</em> spermatogonial cell cryopreservation, aiming to generate a spermatogonia cryobank for future conservation efforts.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141726926","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 predictive value of monocyte-related inflammatory factors for recurrence of atrial fibrillation after cryoablation","authors":"Maimaiti Aimaitijiang ,&nbsp;Aisikaer Gulisitan ,&nbsp;Zhengyan Zhai ,&nbsp;Aili Atawula ,&nbsp;Chunying Jiang","doi":"10.1016/j.cryobiol.2024.104945","DOIUrl":"10.1016/j.cryobiol.2024.104945","url":null,"abstract":"<div><p>Our objective was to investigate the predictive value of monocyte-related inflammatory factors, including monocyte to high-density lipoprotein cholesterol ratio (MHR) and monocyte to lymphocyte ratio (MLR), for the recurrence of atrial fibrillation (AF) after cryoablation in AF patients. The 570 patients who underwent cryoablation were divided into AF recurrence group and non-recurrence group based on follow-up results. The multivariable logistic regression analysis was used to evaluate the effect of MHR and MLR on AF patients. The AF-free survival status of patients was tested by Kaplan-Meier method. ROC analysis was performed to assess the predictive value of MHR and MLR for post-ablation recurrence of AF. A total of 113 (19.8 %) patients relapsed, while 457 patients (80.2 %) had no AF recurrence during follow up. Patients with AF recurrence had higher MHR values (0.37 ± 0.14 vs. 0.33 ± 0.14; <em>P</em> = 0.004) and higher MLR values (0.49 ± 0.32 vs. 0.18 ± 0.07; <em>P</em> &lt; 0.001) compared to those without AF recurrence. MHR≥0.34 combined with MLR≥0.24 (HR = 9.979, 95 % CI: 6.070–16.407, <em>P</em> &lt; 0.001) was an independent factor for predicting AF recurrence after cryoablation in patients by logistic regression analysis. The ROC analysis showed that the AUC for the combination of the MHR and MLR variables was 0.974 (95 % CI: 0.962–0.985) and had the highest diagnostic sensitivity (97.4 %). Elevated baseline values of the monocyte-related inflammatory factors, MHR and MLR, have a certain predictive value for increased AF recurrence after cryoablation.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141757626","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":"l-Proline and GelMA hydrogel complex：An efficient antifreeze system for cell cryopreservation","authors":"Xin Li ,&nbsp;Yukun Cao ,&nbsp;Chenxi Liu ,&nbsp;Jia Tan ,&nbsp;Xinli Zhou","doi":"10.1016/j.cryobiol.2024.104942","DOIUrl":"10.1016/j.cryobiol.2024.104942","url":null,"abstract":"<div><p>Cryopreservation of biological samples is an important technology for expanding their applications in the biomedical field. However, the quality and functionality of samples after rewarming are limited by the toxicity of commonly used cryoprotectant agents (CPAs). Here, we developed a novel preservation system by combining the natural amino acid <span>l</span>-proline (L-Pro) with gelatin methacryloyl (GelMA) hydrogels. Compared with dimethyl sulfoxide (DMSO), L-Pro and GelMA demonstrated excellent biocompatibility when co-culturing with cells. Cryopreservation procedures were optimized using 3T3 as model cells. The results showed that rapid cooling was the most suitable cooling procedure for L-Pro and GelMA among the three cooling procedures. Co-culturing with cells for 3 h before cryopreservation, 6 % L-Pro +7 % GelMA had the highest survival rate, reaching up to 80 %. Differential Scanning Calorimetry (DSC) analysis showed that 6 % L-Pro + 7 % GelMA lowered the freezing point of the solution to −4.2 °C and increased the unfrozen water content to 20 %. To the best of our knowledge, this is the first report of cell cryopreservation using a combination of L-Pro and GelMA hydrogels, which provides a new strategy for improving cell cryopreservation.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-07-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141733659","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":"Analysis of cryopreservation media thermophysical characteristics after ultra-rapid cooling through differential scanning calorimetry","authors":"","doi":"10.1016/j.cryobiol.2024.104939","DOIUrl":"10.1016/j.cryobiol.2024.104939","url":null,"abstract":"<div><p>Cryoprotective agents play a critical role in minimizing cell damage caused by ice formation during cryopreservation. However, high concentrations of CPAs are toxic to cells and tissues. Required concentrations of CPAs can be reduced by utilizing higher cooling and warming rates, but insight into the thermophysical properties of biological solutions in the vitrification method is necessary for the development of cryopreservation protocols. Most studies on thermophysical properties under ultra-rapid cooling conditions have been qualitatively based on visualization. Differential scanning calorimetry methods are ideal for studying the behavior of biomaterials in various freezing conditions quantitatively and accurately, though previous studies have been predominantly restricted to slower cooling rates. Here, we developed an ultra-rapid cooling method for DSC that can achieve minimal cooling rates exceeding 2000 °C/min. We investigated the thermophysical vitrification behavior of ternary solutions of phosphate buffer saline (1X), dimethyl sulfoxide or glycerol and ice blocking polymers (X-1000 or Z-1000). We quantified the impact of solute concentration on ice crystal formation during rapid cooling. Our findings support the expectation that increasing the solute concentration reduces the amount of ice formation, including devitrification. Devitrification increases from 0 % to 40 % (v/v) Me₂SO and then reduces significantly. The relative amounts of devitrification to the total ice formation are 0 %, 60 %, 0 % in 20 %, 40 %, 60 % (v/v) Me₂SO, and 2 %, 48 %, 49 % in 20 %, 40 %, 60 % (v/v) glycerol, respectively. The results suggest that at low concentrations, such as below 20 % (v/v) for Me₂SO or glycerol, increasing the warming rate after ultra-rapid freezing is not essential to eliminate devitrification. Furthermore, ice blocking polymers do not reduce ice formation substantially and cannot eliminate devitrification under ultra-rapid cooling conditions. In conclusion, our results provide insights into the impact of solute concentration on ice formation and devitrification during rapid cooling, which can be practical for optimizing cryopreservation protocols.</p></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":null,"pages":null},"PeriodicalIF":2.3,"publicationDate":"2024-07-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S0011224024000944/pdfft?md5=c51b3a4e80ef740707a50c239efb592e&pid=1-s2.0-S0011224024000944-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141544687","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}