Cryo letters最新文献

{"title":"Exposure of carrot seeds to cryopreservation increases root weight and decreases levels of cell wall-linked phenolics.","authors":"Y Entensa, G Lorente, L Perez-Bonachea, J R Ynchausti, J Martinez, B E Zevallos-Bravo, B Companioni, E Hajari, Y Acosta, H W Pritchard, J C Lorenzo","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Background: </strong>The long-term preservation of plant germplasm is critical to ensure a pool of genetic diversity for future breeding efforts and for conservation management.Although it is important to assess whether the biochemical properties and vigour of the germplasm remain unchanged.</p><p><strong>Objective: </strong>To investigate the potential effects of cryopreservation in liquid nitrogen (LN) as a strategy for conservation of carrot.</p><p><strong>Results: </strong>Seeds were exposed to cryogenic temperatures and growth and development was monitored in Petri dishes (for up to 14 days) and in a pot trial (90 days). By day 7, 53% control seeds had germinated compared with 70% seeds exposed to LN. Biochemical attributes were also assessed. Initial differences were observed in the growth rate of plants, where plants originating from seeds exposed to LN emerged faster than those originating from control seeds. However, at the end of the pot trial, differences were only observed in belowground biomass (i.e., mass of carrots). Biochemical evaluations showed that carotenoids and cell wall-linked phenolics were elevated in carrots produced from seeds exposed to LN.</p><p><strong>Conclusion: </strong>Overall, the results show that cryopreservation can be used as a viable strategy for long-term preservation of carrot germplasm. https://doi.org/10.54680/fr25210110812.</p>","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"46 2","pages":"135-142"},"PeriodicalIF":1.0,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143633683","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ultrastructure assessment of oocyte from mouse with polycystic ovary syndrome following cryopreservation.","authors":"S Afzalnia, F Ghasemian, H S Maryan, T Mirzapour","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Background: </strong>The polycystic ovary syndrome (PCOS) is a substantial obstacle to female fertility due to ovulation inhibition. Oocyte cryopreservation is crucial for preserving fertility in women with fertility-compromising disorders such as PCOS.</p><p><strong>Objective: </strong>In this study, the ultrastructural damages of oocytes were evaluated following freezing from PCOS mouse model.</p><p><strong>Materials and methods: </strong>This experimental study was conducted on 30 adult NMRI mouse. The study comprised three groups: 1) unfrozen PCOS oocytes; 2) vitrified-thawed control oocytes; and 3) vitrified-thawed PCOS oocytes. Transmission electron microscopy was employed for ultrastructure examination across all groups. Moreover, the expression of apoptotic genes, including BAX and Bcl2, was assessed using real time-quantitative polymerase chain reaction (RT-PCR).</p><p><strong>Results: </strong>The oocyte cryopreservation process had a high impact on the destruction of ooplasm cortical granules, Golgi complexes and mitochondria in vitrified-thawed PCOS oocytes compared to the other groups. In PCOS oocytes, particularly those that were vitrified-thawed, there was a notable increase in vacuolation, with a higher abundance of larger and more numerous vacuoles observed compared to the control group. The vitrified-thawed PCOS group also exhibited a notable increase in the expression of the apoptotic gene compared to the other groups (p < 0.05).</p><p><strong>Conclusion: </strong>A precise evaluation of oocyte cryopreservation is imperative for improving this technique and for producing high-quality oocytes with enhanced fertility potential. This study contributes valuable insights into understanding the intricate relationship between PCOS, cryopreservation and oocyte quality. https://doi.org/10.54680/fr25210110412.</p>","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"46 2","pages":"116-125"},"PeriodicalIF":1.0,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143633705","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Fortification of semen extender with mifepristone improves the cryo-survival of cattle spermatozoa.","authors":"A Kumar, V Sachan, M Anand, A Kumar, N Chaudhary, G Singh, M Kumar, J K Agrawal, A Saxena","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Background: </strong>Progesterone, which is present in the semen extender as a component of egg yolk is a potential inducer of capacitation in spermatozoa during cryopreservation. An anti-progesterone component in the extender may protect the spermatozoa from being capacitated and pre-acrosome reacted during cryopreservation. It may lead to better quality of post-thaw sperm population for improved conception.</p><p><strong>Objective: </strong>To investigate the effect of mifepristone on the cryo-survivability of cattle spermatozoa.</p><p><strong>Materials and methods: </strong>Thirty two semen ejaculates were collected from four Sahiwal bulls and divided into three fractions. These fractions were extended with egg yolk-based TRIS extender supplemented with different concentrations of mifepristone (0, 10 and 20 uM) and subjected to cryopreservation. Cryopreserved semen samples were thawed and evaluated for spermatozoa motion parameters (CASA), viability (flow cytometer), hypo-osmotic swelling test (HOST) responsiveness, capacitation status (CTC), acrosome reaction (flow cytometer) and intracellular calcium ion concentrations (flow cytometer).</p><p><strong>Results: </strong>There was no definitive effect of mifepristone on sperm motility and kinematics. However, the semen samples which were treated with mifepristone showed significantly higher spermatozoa viability and HOST responsiveness. Mifepristone also protected spermatozoa from being cryo-capacitated during the preservation process. Higher percentages of uncapacitated and acrosome intact spermatozoa were found at the post-thaw stage in comparison to the untreated group. Mifepristone-treated groups showed fewer spermatozoa with high intracellular calcium levels.</p><p><strong>Conclusion: </strong>A 10 uM concentration of mifepristone has better potential to protect the spermatozoa from progesterone-induced cryo-capacitation and premature acrosome reaction during cryopreservation. https://doi.org/10.54680/fr25210110612.</p>","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"46 2","pages":"126-134"},"PeriodicalIF":1.0,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143633687","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Verification of the decrease in cell recovery after freezing and thawing due to suboptimal shipping using nine cancer cell lines and the differences in impacts between the cell lines.","authors":"M Sato, Y Nakata, M Noguchi, S Araki, Y Satsuo","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Background: </strong>The impacts of suboptimal shipping conditions during transport on cell viability, recovery, and function of cryopreserved samples, have not been well studied.</p><p><strong>Objective: </strong>The impacts of suboptimal shipping on viability and recovery after the freezing and thawing were investigated using nine cancer cell lines, with particular reference to the approximate level of exposure temperature and exposure time at which adverse effects occur, and whether there are differences in sensitivity between cell types.</p><p><strong>Materials and methods: </strong>The adverse effects of any set of suboptimal shipping conditions (-80 degree C for 7 d, -65 degree C or -50 degree C for 1, 3, and 7 d) on nine cancer cell lines (CHO-K1, COS-1, HeLa, HepG2, HL-60, Jurkat, MCF7, MDCK, 293T) were compared with data obtained during storage in liquid nitrogen.</p><p><strong>Results: </strong>No statistically significant decrease in viability was observed in seven of the nine cell lines after freezing and thawing. On the other hand, a statistically significant decrease in the cell recovery was observed after 2 d post freezing and thawing in the nine cell lines, except CHO-K1 at higher exposure temperatures and longer exposure times. Visualization of the adverse effects on the cell lines using a heat map showed that the impacts tended to be more pronounced under the condition of exposure at -50 degree C for three or more days.</p><p><strong>Conclusion: </strong>These results will contribute to the development of standardized protocols and best practices for the optimal shipping of frozen animal cells. https://doi.org/10.54680/fr25210110212.</p>","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"46 2","pages":"108-115"},"PeriodicalIF":1.0,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143633707","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Strategies used by organisms to survive very cold climates - student's guide.","authors":"S P Chesterfield, H V Joyce, T D Fooks, P W Wilson","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>We briefly examine how cold-hardiness in general, including freeze-tolerance, freeze-avoidance and dehydration strategies allow survival in cold climates, through the eyes of some specific insects and fish. Strategies do vary with geography and latitude, even between two types of insects living in the same area. We look at ice nucleation proteins to enhance freezing and antifreeze proteins to help avoid ice formation or, in some cases, to hinder what is known as recrystallization, as a frozen organism thaws. https://doi.org/10.54680/fr25210110112.</p>","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"46 2","pages":"67-73"},"PeriodicalIF":1.0,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143633688","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Formation of an outer glassy shell hinders the drying of trehalose droplets.","authors":"G Pei, Y Zhang, P Zhou, C Gao","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Background: </strong>Airflow drying of mammalian cells has not been successful yet, with one obstacle being the improper drying time of cells-laden trehalose droplets.</p><p><strong>Objective: </strong>To evaluate the major factors affecting the drying kinetics of aqueous trehalose droplets.</p><p><strong>Materials and methods: </strong>A numerical analysis was applied to the evaporation behavior of aqueous trehalose droplets for the preservation of biomaterials. Factors such as convection caused by droplet contraction, vapor diffusion, and Stefan flow around droplets were taken into account in the analysis.</p><p><strong>Results: </strong>Reducing the size and initial concentration of the droplets helps to achieve rapid drying of droplets. Forced convection can effectively enhance the initial drying rate of droplets, which may mitigate cell damage caused by hypertonic solutions. Upon drying, however, the rapid formation of an outer glassy shell inhibits the further drying of trehalose droplets.</p><p><strong>Conclusion: </strong>Simple enhancement of convection does not facilitate complete droplet vitrification due to the formation of an outer glassy shell that prevents water movement. https://doi.org/10.54680/fr25210110712.</p>","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"46 2","pages":"92-97"},"PeriodicalIF":1.0,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143633685","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Production and cryopreservation of 3d cultures.","authors":"N Moisieieva, O Gorina, A Moisieiev, O Prokopiuk","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Three-dimensional (3D) culture systems, which include spheroids (SPs), provide a unique platform for studying complex biological processes in vivo and for enhancing the capabilities of in vitro test systems. Their uniqueness lies in the 3D organization of cells and in the reproduction of complex intercellular interactions, similar to those in native tissues and organs. These mini-organs can be used for fundamental research, tissue-engineering constructs, development of preclinical models for testing pharmacological drugs, etc. Important and current issues regarding SPs involve improving methods for their production and cryopreservation. Solving these issues will expand the range and effectiveness of their use in tissue engineering. Here, we describe the authors' research and experience on factors influencing the formation of SPs, which can enhance the understanding of their correct application and standardization. A crucial aspect of this review is the information on applying theoretical approaches based on physico-mathematical calculations to improve the quality of existing cryopreservation protocols for SPs. https://doi.org/10.54680/fr25110110112.</p>","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"46 1","pages":"1-13"},"PeriodicalIF":1.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143633722","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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 cryolipolysis on subcutaneous adipose tissue of adult women: immunohistochemical analysis.","authors":"C Rt Palauro, P S Palmeira Silva Daumas, E M Carreiro, F Pimentel de Almeida, I L Dias, P F Meyer","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Background: </strong>The skin, the largest organ in the human body, is composed of complex layers that include subcutaneous adipose tissue. Understanding the characteristics of this skin structure is essential to optimize therapeutic interventions, such as cryolipolysis, aiming for more effective and personalized results.</p><p><strong>Objective: </strong>To evaluate the immunohistochemical effects of skin tissue in adult women undergoing cryolipolysis.</p><p><strong>Materials and methods: </strong>We carried out an experimental and blind study with immunohistochemical analysis in women with localized abdominal fat, categorized based on the constitution of the skin as flaccid or firm according to the Investigator Assessment Skin Laxity Scoring System scale. Participants were randomized before undergoing the cryolipolysis procedure. Forty-five days after the procedure, they underwent abdominoplasty, with collection of biological material. We evaluated the inflammatory markers EBF-1, TNF-alpha, and CD68, as well as Caspase 3, cleaved Caspase 3, apoptotic BCL2, Ki-67 for fibroblast proliferation, and FIS1 for mitochondrial proliferation.</p><p><strong>Results: </strong>Six women were included, divided into two groups; three women with loose and three with firm skin. We observed that after cryolipolysis, the group with flaccid skin showed higher expression of the Casp3, TNF-alpha, BCL2, and FIS1 markers compared to those with firm skin.</p><p><strong>Conclusion: </strong>Cryolipolysis may act differently according to tissue morphology, suggesting that its apoptotic response is more pronounced in the group with flaccid skin. https://doi.org/10.54680/fr25110110212.</p>","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"46 1","pages":"41-46"},"PeriodicalIF":1.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143633581","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"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 carnitine on Hariana bull spermatozoa function after cryopreservation.","authors":"B K Yadav, J K Agrawal, K Gangwar, D K Swain, B Yadav, A Kumar, V Sachan, M Anand, B Kumar, A Saxena","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Background: </strong>Carnitine reduces reactive oxygen species-induced apoptosis and DNA fragmentation through its antioxidant effect.</p><p><strong>Objective: </strong>To investigate the effect of carnitine on capacitation, mitochondrial activity, acrosomal integrity, reactive oxygen species (ROS), membrane fluidity, and DNA fragmentation during the cryopreservation of Hariana bull spermatozoa.</p><p><strong>Materials and methods: </strong>Thirty-two semen ejaculates were obtained using artificial vagina (AV) from four seemingly healthy Hariana bulls. Following dilution, the diluted semen samples were split into four aliquots: Group I, the control, included no carnitine; Groups II, III, and IV were the aliquots that contained carnitine supplements of 2.5, 5, and 10 mM, respectively. These four diluted semen samples were then processed immediately for freezing and equilibration.</p><p><strong>Results: </strong>Regarding post-thaw sperm parameters, such as viability, motility, velocity parameters, capacitation status, mitochondrial activity, acrosomal integrity, reactive oxygen species (ROS), membrane fluidity, and DNA fragmentation, Groups II and III, containing 2.5 mM and 5 mM carnitine respectively, had significantly (P < 0.05) improved parameters compared to the Group I (control). At 5 mM, there was a substantial (P < 0.05) decrease in early apoptotic-like alterations in sperm cells, accompanied by a greater population of sperm cells with high mitochondrial membrane potential.</p><p><strong>Conclusion: </strong>Carnitine has been shown to have cryoprotective properties in semen extenders. For improved post-thaw sperm quality, carnitine may be added to a Hariana bull semen extender at a dose of 5 mM. https://doi.org/10.54680/fr25110110712.</p>","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"46 1","pages":"31-40"},"PeriodicalIF":1.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143633519","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Non-permeable cryoprotectants' influence on fibroblast slow freezing in six-banded armadillo.","authors":"D P Fernandes, E A Praxedes, J Vitor da Silva Viana, M Valeria de Oliveira Santos, C I Alves Freitas, A F Pereira","doi":"","DOIUrl":"","url":null,"abstract":"<p><strong>Background: </strong>There is a crucial need to develop appropriate cryopreservation solutions so that somatic resource biobanks of wildlife can be established.</p><p><strong>Objective: </strong>Here, we propose a cryopreservation protocol to optimize the preservation of skin-derived fibroblasts from six-banded armadillos (Euphractus sexcinctus Linnaeus, 1758) by comparing different concentrations of fetal bovine serum (FBS) in the absence or presence of sucrose as non-permeable cryoprotectants.</p><p><strong>Materials and methods: </strong>Cells were cryopreserved by slow freezing with different solutions containing Dulbecco's modified Eagle's medium (DMEM) with 10% dimethyl sulfoxide (DMSO), varying concentrations of FBS (10, 20 and 40%) without or with 0.2 M sucrose, totaling six comparison groups. Cells not subjected to cryopreservation were used as a control. Cells were evaluated for morphological characteristics, viability, metabolism, apoptosis levels, proliferative activity and mitochondrial membrane potential.</p><p><strong>Results: </strong>Cells maintained similar fusiform morphology and demonstrated high viability (> 90%) before and after cryopreservation in all groups. Cryopreserved cells with 10 and 40% of FBS without sucrose showed lower metabolism, but, when sucrose was added, this parameter was maintained as in the control group. This effect was not observed in the 20% FBS groups in the absence or presence of sucrose, with viability similar to that of the non-cryopreserved group. The addition of sucrose maintained apoptosis levels, while the 20 and 40% FBS without sucrose groups showed alterations in viable, early apoptosis and necrosis stages. Nevertheless, all cryopreserved groups showed lower proliferative activity with a higher population doubling time (16.2-19.9 h) than the non-cryopreserved group (15.2 h). Finally, the 20% FBS groups, in the absence or presence of sucrose, maintained the mitochondrial membrane potential.</p><p><strong>Conclusion: </strong>We demonstrated that 20% FBS with sucrose was the most suitable cryopreservation solution for six-banded armadillo skin-derived fibroblast lines, promoting high cell survival after thawing. https://doi.org/10.54680/fr25110110412.</p>","PeriodicalId":10937,"journal":{"name":"Cryo letters","volume":"46 1","pages":"47-56"},"PeriodicalIF":1.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143633641","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}