Cryobiology最新文献

{"title":"Effect of long-term storage on the quality of cryopreserved sperm of the Pangasius nasutus (Bleeker, 1863)","authors":"Nurizzati Idris ,&nbsp;Donald Torsabo ,&nbsp;Muhammad Yazed Abduh ,&nbsp;Ambok Bolong Abol-Munafi ,&nbsp;Noordiyana Mat Noordin ,&nbsp;Ivan Chong Chu Koh","doi":"10.1016/j.cryobiol.2025.105219","DOIUrl":"10.1016/j.cryobiol.2025.105219","url":null,"abstract":"<div><div>In the present study, we investigated the effects of storage for 3, 6, 9 and 12 months on cryopreserved sperm of <em>P. nasutus</em>, in 10 % MeOH as a cryoprotectant with 90 % 0.9 % NaCl as an extender. Sperm quality on motility, fertilization, hatching, and deformity rates were investigated using 9-months cryopreserved sperm. Determination on the effects of different sperm to egg ratios also was evaluated using 1-year cryopreserved sperm. Post-thaw motility (PTM) of cryopreserved sperm within storage period was significantly lower compared to the initial motility of fresh sperm (51.67 ± 2.4 %). However, there was no significant change of PTM during 12 months of storage (3 months, 33.3 ± 2.33 %; 6 months, 32.0 ± 2.52; 9 months, 32.67 ± 1.76; 12 months, 33.33 ± 1.67). In addition, the motility duration was unaffected by storage as there was no significant difference compared to fresh samples. Besides that, there were no significant differences in motility, fertilization and deformity rates for fresh and 9-months cryopreserved sperm except for hatching rate. Samples stored for 1 year resulted in high fertilization across all sperm to egg ratios, showing no difference compared to fresh sperm. Cryopreserved sperm exhibited a significant higher hatching rate (9.24 ± 4.68 %) at the sperm to egg ratio of 300,000:1 compared to 150,000:1 (3.44 ± 1.7 %). However, no significant difference in deformity rates for fresh and cryopreserved at sperm to egg ratio of 300,000:1. The newly developed cryopreservation protocol using methanol successfully preserved sperm quality, maintaining fertilization rates comparable to fresh sperm. This highlights its potential application in sustainable aquaculture and conservation strategies for <em>Pangasius nasutus</em>.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105219"},"PeriodicalIF":2.3,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143593992","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":"Potential application of acid-form sophorolipids in cell cryopreservation","authors":"Thi Nhu Trang Nguyen,&nbsp;Yuki Saito,&nbsp;Motoki Tatsumi,&nbsp;Masashi Yamamoto,&nbsp;Yoshihiko Hirata","doi":"10.1016/j.cryobiol.2025.105228","DOIUrl":"10.1016/j.cryobiol.2025.105228","url":null,"abstract":"<div><div>In cryopreservation, which is an important process for the long-term storage and transport of cells, cryopreservation solutions containing cryoprotectants are usually used to protect cells from damage. However, most cryoprotectants have non-negligible cytotoxicity and side effects that greatly limit their applications in clinical use. This has led to an increased need for more biocompatible cryoprotectants, and interest in bioinspired cryoprotectants is increasing. In this study, we have discovered the potential of using natural acid-form sophorolipids (aSL) as a cryoprotectant, which suppresses ice formation and reduces osmotic stress. The solution containing aSL showed the smallest size of ice crystals compared to the solutions containing other cryoprotectants, such as dimethyl sulfoxide (Me2SO), glycerol (GL), ethylene glycol (EG), and propylene glycol (PG). By adding aSL to a hypertonic culture medium, cell viability was significantly improved. This finding suggests an opportunity to develop low-toxicity and efficient reagents for cell cryopreservation in the future.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105228"},"PeriodicalIF":2.3,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143593993","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 vitrification on in vitro developmental competence of rat testicular tissue","authors":"Mina Sharbatoghli ,&nbsp;Samira Hajiaghalou ,&nbsp;Nafiseh Sadat Deheshkar Gooneh Farahani ,&nbsp;Samaneh Aghajanpour ,&nbsp;Abdolhossein Shahverdi ,&nbsp;Mojtaba Rezazadeh Valojerdi ,&nbsp;Bita Ebrahimi","doi":"10.1016/j.cryobiol.2025.105213","DOIUrl":"10.1016/j.cryobiol.2025.105213","url":null,"abstract":"<div><div>Cryopreservation of testicular tissue has been proposed as a potential technique for preserving fertility in pre-pubertal boys with various malignancies. The present study aimed to compare the effects of two vitrification techniques—solid surface vitrification (SSV) and needle-immersed vitrification (NIV)—on the integrity, development, cell viability, and apoptosis of rat testicular tissue. Testes from 4-week-old Wistar rats underwent a two-step vitrification process. Tissue pieces were allocated to either the SSV or NIV group. Equilibration involved a solution containing 7.5 % dimethyl sulfoxide (DMSO) and 7.5 % ethylene glycol (EG), followed by a vitrification solution with 0.07 mol/L sucrose, 15 % DMSO, and 15 % EG. The optimal protocol was determined after vitrification using either the NIV or SSV technique. Samples from the control and selected vitrification (SSV) groups were cultured for 3 weeks. Tissue integrity, cell viability, apoptosis, and gene expression were evaluated using hematoxylin and eosin staining, trypan blue staining, annexin V-PI staining, and real-time PCR. Morphological changes were more pronounced in the NIV group compared to the SSV group (P &lt; 0.05). Although the percentage of viable cells did not significantly differ between the NIV and SSV groups, it was slightly higher in the SSV group. Thus, SSV was identified as the optimum vitrification method. Real-time PCR analysis revealed altered gene expression: spermatogonial-related genes (<em>Lrp4</em>, <em>Egr3</em>, <em>Nanos</em>, <em>Gfra1</em>, <em>C-kit</em>, and <em>Sohlh1</em>) were significantly decreased in the SSV group, while somatic-cells-related genes (<em>Gdnf</em>, <em>Csf1</em>, and <em>Fgf2</em>) were higher. Overall, SSV appears suitable for rat testis tissue vitrification, although it induces some molecular changes. Optimization of the culture medium is essential to support successful spermatogenesis.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105213"},"PeriodicalIF":2.3,"publicationDate":"2025-03-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143425143","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":"Long term stability of preservative-free and lyophilized PRGF eye drops stored at different temperature conditions: in vitro comparative study","authors":"Eduardo Anitua ,&nbsp;María de la Fuente ,&nbsp;Mohammad Hamdan Alkhraisat","doi":"10.1016/j.cryobiol.2025.105214","DOIUrl":"10.1016/j.cryobiol.2025.105214","url":null,"abstract":"<div><div>Long-term stability of blood-derived eye drops is required to adapt the use of this treatment to prolonged clinical treatments and more green pharmacy. This study has been designed to assess the long-term storage life of freeze-dried plasma rich in growth factors (PRGF) eye drops, maintaining their biological content and activity. Thus, blood from five healthy donors was extracted and centrifuged for obtaining PRGF. The obtained PRGF eye-drops after platelet activation were freeze-stored or were lyophilized and then stored for 18 and 24 months at room temperature (RT) or at + 5 °C. Growth factor content and proliferative potential of PRGF eye drops on primary human keratocytes (HK) was evaluated at each storage time and condition. All growth factors maintained their levels at each time and storage condition. No differences were observed on the proliferative activity of keratocytes after treatment with freeze-dried PRGF eye-drops stored at RT or +5 °C for 18 or 24 months in comparison with fresh samples. No microbial contamination was observed in any of the PRGF eye-drops. Accepting the limitations of this study, it is observed that freeze-dried PRGF eye drops retain both key growth factors and biological activity when stored at room temperature or +5 °C for up to 18–24 months.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105214"},"PeriodicalIF":2.3,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143432687","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 “Cryopreservation of porcine skin-derived stem cells using melatonin or trehalose maintains their ability to self-renew and differentiate” [Cryobiology 107 (2022) 23–34]","authors":"Jia-Dong Sun ,&nbsp;Yu Sun ,&nbsp;Tian Qiao ,&nbsp;Shu-Er Zhang ,&nbsp;Paul W. Dyce ,&nbsp;Yuan-Wei Geng ,&nbsp;Ping Wang ,&nbsp;Wei Ge ,&nbsp;Wei Shen ,&nbsp;Shun-Feng Cheng","doi":"10.1016/j.cryobiol.2025.105201","DOIUrl":"10.1016/j.cryobiol.2025.105201","url":null,"abstract":"","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"118 ","pages":"Article 105201"},"PeriodicalIF":2.3,"publicationDate":"2025-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143028151","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":"Effect of resveratrol on post-thaw motility, kinematics, structural parameters and antioxidant/oxidant status of Kamori buck spermatozoa","authors":"Tarique Hussain ,&nbsp;Muhammad Hammad Fayyaz ,&nbsp;Amjad Hameed ,&nbsp;Syed Murtaz Hassan Andrabi ,&nbsp;Rehana Kausar ,&nbsp;Yasin Mubashir ,&nbsp;Iqra Batool ,&nbsp;Muhammad Shahzad ,&nbsp;Ali Dogan Omur","doi":"10.1016/j.cryobiol.2025.105202","DOIUrl":"10.1016/j.cryobiol.2025.105202","url":null,"abstract":"<div><div>Resveratrol is a polyphenol compound showing strong antioxidant properties. It is believed that semen cryopreservation causes significant sperm losses which eventually affects sperm quality. Improving antioxidant status of semen may reduce this damage and enhance sperm fertilizing potential. This study investigated the resveratrol treated freeze-thawing sperm motion, kinetics, structural parameters and antioxidant/oxidant status of Kamori buck spermatozoa. Thirty-two ejaculates from 4 fertile Kamori bucks were processed in tris-citric-glyercol-egg yolk (TCG-EY) based with varying concentrations of resveratrol (0, 10, 20, 40 and 50 μM). Over 75 % sperm motility were pooled and frozen in liquid nitrogen. The results unveiled that adding resveratrol at 40 and 50 μM concentration significantly enhanced (<em>P</em> &lt; 0.05) total motility (progressive motility, rapid velocity, and average path velocity, straight line velocity, curvilinear velocity, amplitude of lateral head displacement, beat cross frequency, straightness and linearity in contrast with control and other groups. Supplementing resveratrol at 40 and 50 μM concentration significantly improved (<em>P</em> &lt; 0.05) functional plasma membrane integrity, acrosome integrity, whereas, all resveratrol groups had same significant (<em>P</em> &lt; 0.05) effect on DNA integrity in response to control group. The 40 and 50 μM resveratrol significantly promoted (P &lt; 0.05) superoxide dismutase (SOD), catalase (CAT), peroxidases (POD), and total antioxidant capacity (TAC) levels while significantly reducing (<em>P</em> &lt; 0.05) the total oxidant status (TOS) and malondialdehyde (MDA) contents as compared to control and other groups, respectively. The principal component analysis (PCA) exhibited samples were present in different clusters except two groups which had partially overlapped. Using hierarchical clustering analysis, two clusters were constructed showing the relationship within the treated groups. The results of spearman correlation coefficient showed that yellow color showed highly positive correlation while turquoise color exhibited highly negative association between sperm variables. Overall, the results concluded that resveratrol at 50 μM performed slightly better results than 40 μM in terms of improving sperm quality parameters.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"118 ","pages":"Article 105202"},"PeriodicalIF":2.3,"publicationDate":"2025-02-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143001673","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":"Is isochoric vitrification feasible? Cycling between modeling, experimental observations, and first principles","authors":"Yoed Rabin,&nbsp;Prem K. Solanki","doi":"10.1016/j.cryobiol.2025.105216","DOIUrl":"10.1016/j.cryobiol.2025.105216","url":null,"abstract":"<div><div>Isochoric cryopreservation is preservation of biological material in a constant-volume container, to benefit from a presumed baroprotective effect. From its inception, the premise for the isochoric approach is that pressure elevation can be achieved by permitting some ice formation in the system. The idea of integrating vitrification into an isochoric chamber followed at a later stage, where vitrification is a century old established approach of ice-free cryopreservation. At face value, the resulting term “isochoric vitrification” encompasses an internal contradiction revolving around ice formation. We have previously shared modeling results pointing to this contradiction and, hence, questioning the feasibility of the process (Cryobiology 2023, 111, 9–15). More recently, Ali et al. (Cryobiology 2024, 116, 104935) claimed to have obtained contradictory experimental evidence while stating that isochoric vitrification is feasible. The objective for this communication is to share our skepticism about this claim with the community of cryobiology.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"118 ","pages":"Article 105216"},"PeriodicalIF":2.3,"publicationDate":"2025-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143467291","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":"Assessment of alternative freezing methods for preservation at −80 °C of radiata pine embryogenic cultures: A six-year study","authors":"K.P. Sandoval,&nbsp;A. Castander-Olarieta,&nbsp;P. Moncaleán,&nbsp;I.A. Montalbán","doi":"10.1016/j.cryobiol.2025.105217","DOIUrl":"10.1016/j.cryobiol.2025.105217","url":null,"abstract":"<div><div>Somatic embryogenesis is an essential component of breeding programs for <em>Pinus radiata</em> aimed at implementing multi-varietal forestry. Coupled with this technique, the long-term cryopreservation of embryogenic cultures is necessary to maintain the viability of the cell lines, but this entails high maintenance costs. In this research we evaluated the application of a protocol for long-term storage at −80 °C in an ultra-low freezer to preserve several radiata pine embryogenic cell lines. Also, we studied the influence of several parameters to optimize the protocol, such as the effect of dimethyl sulfoxide cryoprotectant solution, the effectiveness of alternative freezing methods, the use of post thawing treatments and the addition of sodium butyrate at maturation stage. We found that the use of dimethyl sulfoxide cryoprotectant enhanced somatic embryo production; slow cooling was the only viable method for preserving embryogenic cell lines at −80 °C and the use of sodium butyrate was not highly effective to improve maturation and germination stages. Moreover, we have regenerated embryogenic cell lines up to their conversion into plants after six years of storage. In line with these findings, the protocol to storage in an ultra-low freezer represents an economical alternative to preserve somatic embryogenic cultures of <em>Pinus radiata</em>.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105217"},"PeriodicalIF":2.3,"publicationDate":"2025-02-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143482475","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":"Procyanidin B2 alleviates damage to mouse testicular tissue after freezing by inhibiting oxidative stress and apoptosis","authors":"Guorui Cao ,&nbsp;Chunyuan Li ,&nbsp;Jian Zhang ,&nbsp;Liwen Deng ,&nbsp;Rong Li ,&nbsp;Changlong Xu","doi":"10.1016/j.cryobiol.2025.105196","DOIUrl":"10.1016/j.cryobiol.2025.105196","url":null,"abstract":"<div><div>For infertile patients who are unable to obtain sperm or prepubertal boys who require radiotherapy, testicular tissue freezing can be used for later transplantation and is a potentially effective method of preserving male fertility. Oxidative stress caused by the freezing process is an important cause of tissue damage. Procyanidin B2 (PCB2) is a polyphenolic natural compound widely distributed in plants that is known for its anti-inflammatory, anticancer, and neuroprotective properties, and its antioxidant capabilities are particularly noteworthy. Research has indicated that PCB2 exerts a protective effect on the reproductive system. However, its specific role in mitigating testicular tissue cryoinjury and the underlying mechanisms remain unclear. This study investigated whether adding PCB2 to a cryoprotective solution of mouse testicular tissue can alleviate the cryoinjury of testicular tissue and its possible mechanism. Our findings revealed that frozen mouse testicular tissue presented decreased cell viability and induced oxidative stress. Conversely, PCB2 effectively mitigated these adverse effects. In addition, PCB2 improved the tubular structural disorganization caused by freezing and increased the expression of proteins related to the junction function of Sertoli cells. Further experiments indicated that PCB2 activated the nuclear respiratory factor 2 (Nrf2)/heme oxygenase 1 (HO-1) antioxidant signaling pathway, increased the activity of downstream antioxidant enzymes, and improved mitochondrial kinetic homeostasis. Additionally, PCB2 ameliorated apoptosis while increasing the expression levels of key enzymes involved in testosterone synthesis. In summary, these results suggest that PCB2 attenuates damage to mouse testicular tissue during freezing by inhibiting oxidative stress and apoptosis.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"118 ","pages":"Article 105196"},"PeriodicalIF":2.3,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142930567","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":"Mitochondrial respiratory analysis of cryopreserved PBMCs isolated from human blood","authors":"C. Payne ,&nbsp;E. Louw ,&nbsp;N. Baines ,&nbsp;B. Botha ,&nbsp;C. Lombard ,&nbsp;B. Allwood ,&nbsp;G. Maarman","doi":"10.1016/j.cryobiol.2025.105212","DOIUrl":"10.1016/j.cryobiol.2025.105212","url":null,"abstract":"<div><div>Mitochondrial bioenergetics of PBMCs have been linked with several factors that contribute to a better understanding of several human diseases. Due to the complex logistics of clinical studies, samples are often cryopreserved for later analysis. Current data on whether cryopreservation negatively affects the mitochondrial function of PBMCs is discrepant. We isolated and cryopreserved peripheral blood mononuclear cells (PBMCs) from human whole blood and tested mitochondrial function using a substrate-uncoupler-inhibitor-titration protocol on the Oroboros instrument. After three months of storage in a cryopreservation medium (at −80 °C), several aspects of mitochondrial bioenergetics were measured. We demonstrate that cryopreservation did not adversely affect mitochondrial parameters (routine, leak, complex-I linked OXPHOS, cytochrome-c response, ETS capacity, the contributions of the N and S-pathways to ETS, ROX, complex-IV activity and mitochondrial coupling). Therefore, after three months of cryopreservation at −80 °C, human PBMC-mitochondria were fully coupled and functional. Therefore, clinical studies may cryopreserve PBMCs for later mitochondrial analyses.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"118 ","pages":"Article 105212"},"PeriodicalIF":2.3,"publicationDate":"2025-02-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143373794","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}