Cryobiology最新文献

{"title":"Ultrasound-based geometric modeling of the human ovary with applications to cryopreservation.","authors":"Rounak K Baheti, Prem K Solanki, Sally Ahmed, Angela Baerwald, Yoed Rabin","doi":"10.1016/j.cryobiol.2024.105187","DOIUrl":"10.1016/j.cryobiol.2024.105187","url":null,"abstract":"<p><p>Successful cryopreservation of the whole ovary outside of the body, while a woman undergoes cancer treatments, may help preserving fertility and regaining hormone balance during recovery. One of the key challenges in whole ovary cryopreservation is adequately loading the organ with cryoprotective agents (CPAs). Another notable challenge in developing the application is the lack of geometric data needed for designing matching thermal protocols. The objective of the current study is twofold: (i) to develop an effective geometric reconstruction method for the ovary, based on transvaginal ultrasound (TVUS) data, and (ii) to perform a pilot study on the thermal effects associated with CPA loading with application to vitrification. This study includes screening of 127 TVUS imaging datasets of ovaries from healthy ovulatory participants, reconstruction of 14 geometric models, and thermally analyzing two representative geometric models of low and high mature follicles-to-organ volume ratios. Results of this study demonstrate that the proposed reconstruction method is faster and more accurate than that facilitated by commercially available software (SonoAVC, GE Healthcare). Two extremes were investigated: (1) complete vitrification of the ovary, and (2) crystallization of mature follicles while the remaining ovarian stroma vitrifies. CPA loading into the mature follicles is considered an outstanding cryopreservation challenge, but with very little impact on long-term fertility preservation. Results of this study suggest that ovarian preservation by vitrification is feasible when sufficient CPA loading is achieved, while identifying the most suitable CPA for the task remains a challenge beyond the scope of the current study.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105187"},"PeriodicalIF":2.3,"publicationDate":"2024-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142827727","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":"An efficient combination of a CPA loading protocol, metal container, and storage method for successful articular cartilage vitrification.","authors":"Mohammadhamed Shahsavari, Ana Paula Ribeiro Rodrigues, Janet A W Elliott, Nadr M Jomha","doi":"10.1016/j.cryobiol.2024.105185","DOIUrl":"10.1016/j.cryobiol.2024.105185","url":null,"abstract":"<p><p>Successful cryopreservation of articular cartilage (AC) depends on a number of variables, including heat transfer, sample packaging for cryogenic storage, cryoprotectant agent (CPA) concentration (toxicity), and CPA permeation into AC. In the first experiment of the present study, we used a combination of our established vitrification protocol (430-min multi-step loading Protocol 8) and the metal Ovarian Tissue Cryosystem (OTC) as a closed vitrification-storage-rewarming container to vitrify 7-mm diameter porcine osteochondral dowels with 2 methods of storage (storage in the OTC with or without a surrounding vitrification solution). In the second experiment, in an attempt to introduce a more reproducible and safe vitrification protocol, we employed our successful Protocol 8 with the OTC as the CPA loading container and a cryobag as the storage container (with or without surrounding vitrification solution). In both experiments, post-warming normalized chondrocyte viabilities were assessed. The results of the first experiment demonstrated that a combination of the established step-wise CPA loading-vitrification-rewarming protocol (Protocol 8) with the metal OTC as the only container through all steps of CPA loading, vitrification, and rewarming produced a positive result. In the second experiment, combination of Protocol 8, the OTC as the CPA loading system and a cryobag as the storage container in both vitrified groups with or without solution resulted in high normalized chondrocyte viabilities (94.63 % and 96.17 %, respectively) post vitrification in 7-mm diameter porcine osteochondral dowels. Overall, this study demonstrated that by employing our established Protocol 8, using the metal OTC as a CPA loading container and a cryobag as the storage method we could achieve a reproducible method for clinical applications involving vitrified articular cartilage.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105185"},"PeriodicalIF":2.3,"publicationDate":"2024-12-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142784412","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":"Glycerol improves the viability of a cryopreserved choanoflagellate.","authors":"Sheel Chandra, Florentine U Rutaganira","doi":"10.1016/j.cryobiol.2024.105183","DOIUrl":"10.1016/j.cryobiol.2024.105183","url":null,"abstract":"<p><p>The colonial choanoflagellate Salpingoeca rosetta is a tractable model system for studying the origins of multicellularity, but long-term storage strategies for this species have not been tested. In this study, we probed each stage of cryopreservation (cooling, long-term storage, recovery) to identify the optimal protocol for recovery of S. rosetta and co-cultured bacterial cells. Dimethyl sulfoxide (Me2SO; commonly referred to as DMSO), the current cryoprotective agent (CPA) standard, proved to be worse than glycerol at comparable concentrations. Samples treated with either CPA at 5 % showed the poorest recovery. Our results identified 15 % glycerol as the most effective CPA for both S. rosetta and Echinicola pacifica. We also determined that ultra-low temperature freezers can be sufficient for short-term storage. We propose 15 % glycerol and liquid phase nitrogen as the standard cryopreservation protocol for S. rosetta cultures and as a starting point for testing long-term storage strategies for other choanoflagellates and heterotrophic protists.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105183"},"PeriodicalIF":2.3,"publicationDate":"2024-12-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142767141","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":"Effects of low temperature stress on cold-resistant substances and gene expression in Spodoptera frugiperda (Lepidoptera: Noctuidae).","authors":"Huawei Ren, Junrui Zhi, Dingyin Li, Wenbo Yue, Li Liu","doi":"10.1016/j.cryobiol.2024.105166","DOIUrl":"10.1016/j.cryobiol.2024.105166","url":null,"abstract":"<p><p>The fall armyworm Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) is a major agricultural pest. Low temperature is an important factor that limits S. frugiperda survival and reproduction. The physiological cold tolerance indexes of S. frugiperda larvae from October 2022 to February 2023 were investigated. The results showed that the cold-resistant substances in S. frugiperda differed significantly among different months. The water content was the lowest in January. Lipids were the highest in December and January, trehalose and protein contents were the highest in December, and glycerol content was the highest in January. The contents of lipid, protein, trehalose, and glycerol were the highest in the 6^th instar larvae, whereas the water content did not differ among the larva stages. The functions of two genes, including endothelial lipase (Eol1) and gastric lipase (Epl1) that were extracted from the transcriptome analysis of S. frugiperda in the cold resistance were assessed. After 24 h of cold stress, the mRNA levels of Eol1 and Epl1 RNA interference were significantly down-regulated by 60.76 % and 82.53 %, and the survival rate of S. frugiperda larvae decreased. Additionally, Eol1 and Epl1 silencing significantly decreased the lipid content and pupal weight of S. frugiperda. The results indicate that S. frugiperda larvae can change the contents in winter to response the cold conditions and the lipids plays an important role in the cold resistance of S. frugiperda.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105166"},"PeriodicalIF":2.3,"publicationDate":"2024-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142779646","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 thermodynamic principles of isochoric freezing pressure-aided supercooling.","authors":"Alan L Maida, Pedro Alejandro Perez, Cristina Bilbao-Sainz, Boris Rubinsky, Anthony N Consiglio","doi":"10.1016/j.cryobiol.2024.105168","DOIUrl":"10.1016/j.cryobiol.2024.105168","url":null,"abstract":"<p><p>This study outlines a method for designing an isochoric (constant volume) system to reduce the supercooling preservation temperature without affecting the likelihood of ice nucleation and without the need for cryoprotective additives. The method involves a multiphase system wherein the biological material is separated from a second aqueous solution by a boundary that transfers pressure and heat but not mass. The pressure within the system is passively increased by the confined growth of ice within the secondary solution. This increased pressure in turn lowers the equilibrium freezing temperature of the biological matter, which may be utilized to lower the preservation temperature while maintaining the same degree of supercooling. For example, using this technique, the supercooling preservation temperature may be lowered from -2 °C to -5 °C without increasing the risk of ice nucleation, by ensuring the freezable phase makes up ∼17 % of the total system volume.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105168"},"PeriodicalIF":2.3,"publicationDate":"2024-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142667306","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":"Sperm cryopreservation and reproductive characteristics in the beauty snake (Elaphe taeniura) as a model for endangered Colubridae.","authors":"P Banchi, F Solanes-Vilanova, M Cesauri, L Spanoghe, K Chiers, A Van Soom, T Hellebuyck, G Rizzoto","doi":"10.1016/j.cryobiol.2024.105184","DOIUrl":"10.1016/j.cryobiol.2024.105184","url":null,"abstract":"<p><p>Sperm collection and cryopreservation are key technologies for assisted reproduction, with post-mortem sperm collection being the main tool to prevent the decline of endangered species. The present study describes post-mortem sperm collection and two cryopreservation protocols in beauty snakes (Elaphe taeniura), as a model for Colubridae. The vasa deferentia of 18 snakes were collected post-mortem and incubated for 30 min at room temperature to retrieve sperm by float up. Afterwards, fresh sperm was analysed, and 11 samples were diluted into either a single or a double-step TEST-egg yolk-based extender to reach a final concentration of 12 % glycerol. Anatomical and histological analyses of the reproductive organs revealed that those on the right side were larger, heavier, and positioned more cranially than the left ones (P = 0.007, P = 0.003, and P = 0.0002, respectively), but sperm parameters did not differ. Successful sperm retrieval was achieved in 61.1 % of snakes, with positive correlations between seminiferous tubules length and body weight (P = 0.0004) and between seminiferous tubules length and body length (P = 0.001). Sperm-producing animals were heavier and longer compared to azoospermic ones (P = 0.04 and P = 0.04 respectively). Specifically, males longer than 130 cm and heavier than 177g have higher reproductive potential. Both cryopreservation protocols yielded comparable results, preserving sperm viability, motility, and morphology. Therefore, a single-step protocol should be preferred due to its technical simplicity. In conclusion, we described for the first time the reproductive features and characteristics of spermatozoa collected post-mortem in beauty snakes. Although the described protocols are effective, further research is warranted to optimize these techniques.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105184"},"PeriodicalIF":2.3,"publicationDate":"2024-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142767146","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":"An efficient droplet-vitrification cryopreservation procedure for high imperatorin-yielding hairy root clones of Urena lobata.","authors":"Dai Minh Cao, Anh Lan Bui, Le Van Bui, Phuong Ngo Diem Quach","doi":"10.1016/j.cryobiol.2024.105186","DOIUrl":"10.1016/j.cryobiol.2024.105186","url":null,"abstract":"<p><p>The valuable anti-cancer and anti-inflammatory secondary metabolite, imperatorin, has been found in the hairy roots (HRs) of Urena lobata. However, an increasing number of problems related to cryo-injury and cryoprotectant toxicity could potentially reduce the quality of root clones, highlighting the need to develop a reliable technique for long-term preservation. Based on the impact of the successive steps of the initial droplet-vitrification procedure employed for cryopreservation of HRs using the histological evaluation of plasmolysis, various selected factors were independently investigated. The maximum plasmolysis was observed after the preculture with 0.5 M sucrose (46.59 %) and the dehydration treatment (48.32 %). In the improved cryopreservation procedure, when prolonged preculture for 72 h in liquid WPM with 0.3 M sucrose and dehydration in the appropriate vitrification solution, which included 30 % (w/v) glycerol and 50 % (w/v) sucrose, for 10 min at 0 °C, the plasmolysis in the two steps was significantly reduced in comparison with the untreated control. The cryopreserved HRs could increase their regeneration to 93.3 % and regenerated root length to 4.65 cm. Their growth and imperatorin production were almost the same as those of untreated controls after three to five subsequent subcultures. Our results have demonstrated that our new approach, which only focuses on the key factors that significantly increase the plasmolysis, modifies their level to fit with the HR cells of U. lobata. The early application of the plasmolysis evaluation method may immediately screen the impact of factors on individual root cells instead of spending time and cost evaluating the recovery of root tips at each step to develop an efficient cryopreservation procedure.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105186"},"PeriodicalIF":2.3,"publicationDate":"2024-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142767132","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":"No synergistic effect of extracellular cryoprotectants with dimethyl sulfoxide in the conservation of northern tiger cat fibroblasts.","authors":"João Vitor da Silva Viana, Lhara Ricarliany Medeiros de Oliveira, Luanna Lorenna Vieira Rodrigues, Yasmin Beatriz França Moura, Antonia Beatriz Mendonça Pereira, Patrícia Vasconcelos Alves, Herlon Victor Rodrigues Silva, Alexsandra Fernandes Pereira","doi":"10.1016/j.cryobiol.2024.105169","DOIUrl":"10.1016/j.cryobiol.2024.105169","url":null,"abstract":"<p><p>The success of somatic cell cryobanks is dependent on establishing reproducible cryopreservation methodologies. We supposed that associated extracellular cryoprotectants (sucrose and L-proline) with 2.5 or 10 % dimethyl sulfoxide (Me₂SO) could guarantee better northern tiger cat cells quality rates after thawing when compared to Me₂SO alone. Therefore, we evaluated the effects of sucrose or L-proline with 2.5 or 10 % Me₂SO on the cryopreservation of northern tiger cat fibroblasts. Somatic cells were also cryopreserved with 2.5 % or 10 % Me₂SO alone. All cells were analyzed for morphology, membrane integrity, proliferative activity, metabolism, apoptosis classification, reactive oxygen species (ROS) levels, and mitochondrial membrane potential (ΔΨm). Regardless of the cryoprotective solution, cryopreservation did not affect morphology, membrane integrity after culture, proliferative activity, and metabolism (P > 0.05). However, immediately after thawing, 2.5 % Me₂SO with L-proline and 10 % Me₂SO promoted higher rates of membrane integrity when compared to the other cryopreserved groups (P < 0.05). Interestingly, cells cryopreserved with 10 % Me₂SO maintained ROS levels similar to non-cryopreserved cells (P > 0.05). However, the percentage of viable cells evaluated by apoptosis classification was reduced when using 10 % Me₂SO with L-proline compared to non-cryopreserved groups (P < 0.05). Additionally, ΔΨm was altered in all cryopreserved groups (P < 0.05). In summary, sucrose and L-proline were less effective in cryopreservation of northern tiger cat fibroblasts in the presence of 2.5 % or 10 % Me₂SO. Also, 10 % Me₂SO appears to be the most suitable cryoprotectant for the formation of cryobanks of this species.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105169"},"PeriodicalIF":2.3,"publicationDate":"2024-12-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142692306","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":"Cryo-conservation of temperate berry crops at ICAR-NBPGR","authors":"Sandhya Gupta ,&nbsp;Narender Negi ,&nbsp;Sheikh M. Sultan","doi":"10.1016/j.cryobiol.2024.105034","DOIUrl":"10.1016/j.cryobiol.2024.105034","url":null,"abstract":"","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"117 ","pages":"Article 105034"},"PeriodicalIF":2.3,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142743137","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 with ice nucleators enables high-throughput screening of T-Cells","authors":"Natalia Gonzalez-Martinez ,&nbsp;Matthew Gibson","doi":"10.1016/j.cryobiol.2024.104992","DOIUrl":"10.1016/j.cryobiol.2024.104992","url":null,"abstract":"","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"117 ","pages":"Article 104992"},"PeriodicalIF":2.3,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142746950","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}