Cryobiology最新文献

{"title":"Effects of alpha-lipoic acid and sildenafil citrate on sperm quality in asthenozoospermic men during freezing-thawing processes.","authors":"Ronak Kohzadi, Ebrahim Cheraghi, Malek Soleimani Mehranjani","doi":"10.1016/j.cryobiol.2024.105163","DOIUrl":"10.1016/j.cryobiol.2024.105163","url":null,"abstract":"<p><p>We aimed to investigate the combined effects of alpha-lipoic acid (ALA) and sildenafil citrate (SC) on sperm quality during freeze-thawing in asthenozoospermic men. We categorized thirty semen samples from asthenozoospermic into five different groups for analysis: Control (fresh), Freeze, Freeze + SC, Freeze + ALA, and Freeze + SC + ALA. We evaluated sperm parameters in each group, including motility, viability, morphology, plasma membrane integrity, DNA fragmentation, mitochondrial membrane potential, protamine deficiency, acrosome integrity, malondialdehyde, tumor necrosis factor-alpha (TNF-α), antioxidants (Superoxide dismutase, Glutathione, Catalase), and gene expression of HSP70 and Bcl-2. The Freeze group showed significant declines in viability, motility, morphology, membrane integrity, antioxidants, and mitochondrial membrane potential, with increases in protamine deficiency, abnormalities, DNA fragmentation, TNF-α, and malondialdehyde compared to control (p = 0.000). These effects were significantly reversed in all treatment groups (p = 0.000). Bcl-2 and HSP70 expression increased significantly in all groups (p = 0.000). Acrosomal integrity decreased significantly (p = 0.000) in the Freeze group compared to controls and in the Freeze + SC group compared to the Freeze group. However, acrosomal integrity significantly increased (p = 0.000) in the Freeze + SC + ALA group compared to the Freeze + SC group and in the Freeze + ALA group compared to other treatments. Our findings indicate that the simultaneous addition of SC and ALA in the freezing media yielded increased sperm quality. This enhancement was evidenced by reductions in oxidative stress, inflammation, and apoptosis, along with partial prevention of premature acrosome reactions induced by SC.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105163"},"PeriodicalIF":2.3,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142616415","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 oxidative stress on the changes of viability of Paeonia lactiflora seeds with different water content before and after cryopreservation.","authors":"Ji Zhe, Guo Jiayi, Huang Jiaqi, Zhu Qinglu, Ren Ruifen","doi":"10.1016/j.cryobiol.2024.105165","DOIUrl":"10.1016/j.cryobiol.2024.105165","url":null,"abstract":"<p><p>Cryopreservation in liquid nitrogen (LN) is an efficient long - term seed preservation strategy and water content is one of the main factors affecting seed viability after cryopreservation. In this study, Paeonia lactiflora seeds with varying water content were used as materials to determine changes in the antioxidant system before and after cryopreservation. When the water content of P. lactiflora seeds was 15.96 %-10.12 %, the viability of P. lactiflora seeds decreased significantly compared with that of the seeds without cryopreservation, but at the water content of 8.14 %-7.56 % there were no significant differences. However, when the water content of P. lactiflora seeds decreased to 6.01 %-5.19 % the viability was slightly increased compared to the seeds without cryopreservation. The content of superoxide anion (O₂^-), hydroxyl radical (·OH), hydrogen peroxide (H₂O₂), malondialdehyde (MDA) and protein carbonyl group (PCO) of seeds with high water content (15.96 %-10.12 %) were significantly increased after cryopreservation. However, superoxide anion (O₂^-), hydroxyl radical (·OH) and hydrogen peroxide (H₂O₂) did not change significantly in seeds with low water content (6.01 %-5.19 %) after cryopreservation, while oxidative stress indexes MDA and PCO decreased. These three substances were significantly negatively correlated with seed viability. In terms of antioxidant substances, the contents of catalase (CAT), ascorbic acid (AsA) and glutathione (GSH) decreased and the activities of glutathione reductase (GR) and dehydroascorbate reductase (DHAR) increased during the cryopreserved process of seeds with varying water content. These changes were significantly correlated with ROS content and the changes of MDA and PCO, among which AsA content, GSH content and CAT activity were positively correlated with seed viability. The changes of GR and DHAR activity were negatively correlated with seed viability. In summary, when the water content of the seeds ranged from 8.14 % to 5.19 %, ROS content did not increase significantly after cryopreservation compared with that of before preservation. The changes of MDA and PCO contents before and after cryopreservation, it was inferred that no obvious oxidative damage occurred in seeds, so the viability of seeds did not decrease compared with that of before cryopreservation. Therefore, the optimum water content of P. lactiflora seeds for cryopreservation is 8.14 %-5.19 %.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105165"},"PeriodicalIF":2.3,"publicationDate":"2024-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142643715","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":"Nanocrystalline cerium dioxide reduces recrystallization in cryopreservation solutions.","authors":"Olena Bobrova, Oksana Falko, Anna Polyakova, Volodymyr Klochkov, Milosh Faltus, Viktor Chizhevskiy","doi":"10.1016/j.cryobiol.2024.105167","DOIUrl":"10.1016/j.cryobiol.2024.105167","url":null,"abstract":"<p><p>Nanocrystalline cerium dioxide is able to protect living cells from oxidative stress under the influence of various stress factors, in particular under the one of low temperatures. This study investigates the phase-structural transformations in aqueous solutions containing CeO₂ nanoparticles (NPs) and their impact on the cryopreservation process. Differential scanning calorimetry and thermomechanical analysis were used to analyse the phase transitions in aqueous suspensions of CeO₂ NPs and aqueous solutions of the cryoprotectant dimethyl sulfoxide (Me₂SO) with CeO₂ NPs. Various concentrations of CeO₂ NPs were tested to observe their effects on the crystallization and melting behaviours. The addition of CeO₂ NPs significantly altered the temperatures and enthalpies of melting and crystallization in water. Low concentrations of CeO₂ NPs promoted crystallization, while higher concentrations inhibited it, reducing supercooling and recrystallization during thawing. In Me₂SO solutions, CeO₂ NPs raised the glass transition temperature and affected the recrystallization process, with higher concentrations leading to more pronounced vitrification and reduced recrystallization. We also investigated the regularities of the effect of CeO₂ NPs on phase transitions in combined cryoprotective media with Ham's F12, fetal bovine serum and Me₂SO, which can be used in future to design the cryopreservation protocols. In the complex media, CeO₂ NPs decreased the metastability and altered eutectic crystallization patterns, indicating potential cryoprotective effects. In conclusion, CeO₂ NPs modulate the thermophysical properties of cryoprotective solutions, enhancing vitrification and reducing recrystallization, which could improve cryopreservation efficiency. Optimizing NP concentrations is crucial for practical applications in cryopreservation.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105167"},"PeriodicalIF":2.3,"publicationDate":"2024-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142667295","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 -2ºC to -5ºC 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":"2024-11-16","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":"KCl enhances the cryoablation-induced antitumor immune response: A hepatocellular carcinoma murine model research","authors":"Mu Chen ,&nbsp;Baolin Liu","doi":"10.1016/j.cryobiol.2024.105164","DOIUrl":"10.1016/j.cryobiol.2024.105164","url":null,"abstract":"<div><div>Cryoablation is a valuable treatment for liver cancer. To investigate the effect of KCl solution on the immunological response post cryoablation, we created a tumor-bearing mice model by subcutaneously implanting Hepal-6 cells in adult Balb/c mice. Subsequently, the mice were randomly assigned to three groups: group A (sham cryoablation), group B (cryoablation), and group C (cryoablation plus KCl solution). Mice were sacrificed on days 0, 7, and 14 post-treatment. Immune cell populations were assessed using flow cytometry. Blood samples were analyzed for serum IL-4, HSP70, and TGF-β1 levels with ELISA assays. Ablated tissues stained with immunohistochemistry were utilized to evaluate Ki67 expression at the margins of the ablation site. Our findings revealed higher HSP70 expression levels in groups B and C compared to group A. Cryoablation triggered an immune response, which was enhanced by KCl. On days 0, 7, and 14, the percentages of CD4⁺ T cells, CD8⁺ T cells, and NK cells in the spleen of group C were significantly increased compared with groups A and B. Additionally, the Th1/Th2 ratio was significantly increased in group C. Serum TGF-β1 expression was elevated after cryoablation, but KCl solution reduced the high TGF-β1 expression after cryoablation and decreased the invasiveness of cancer cells. Finally, the proliferative activity of untreated tumor tissue was significantly reduced in group C compared to groups A and B. In summary, Cryoablation triggered a systemic immune response in tumor-bearing mice, which was further boosted by combining cryoablation with a KCl solution.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"117 ","pages":"Article 105164"},"PeriodicalIF":2.3,"publicationDate":"2024-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142616416","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":"Biopsy vitrification: New tool for endometrial tissue cryopreservation for research applications","authors":"Merli Saare ,&nbsp;Monika Wróbel ,&nbsp;Yanyu Jiang ,&nbsp;Kenny A. Rodriguez-Wallberg ,&nbsp;Arturo Reyes Palomares ,&nbsp;Keiu Kask ,&nbsp;Aive Kalinina ,&nbsp;Apostol Apostolov ,&nbsp;Ave Minajeva ,&nbsp;Kristina Kiisholts ,&nbsp;Amruta D.S. Pathare ,&nbsp;Piotr Laudański ,&nbsp;Maire Peters ,&nbsp;Andres Salumets","doi":"10.1016/j.cryobiol.2024.105161","DOIUrl":"10.1016/j.cryobiol.2024.105161","url":null,"abstract":"<div><div>Patient-derived endometrial biopsies serve as a crucial source for molecular studies, highlighting the necessity for tissue cryopreservation methods that preserve cell viability and tissue morphology with minimal to no impact. The passive slow freezing (PSF) protocol has demonstrated efficacy for cryopreserving endometrial biopsies, allowing for the subsequent isolation of viable epithelial and stromal cells. Vitrification (VT) enables the avoidance of ice crystal formation and could therefore potentially prevent mechanical injury to tissues. In this study, PSF and VT techniques were applied to endometrial biopsies, and the effects of cryopreservation on tissue samples were evaluated using traditional histology. In addition, transmission electron microscopy (TEM), gene expression profiling analyses, the viability of endometrial cells, and the ability to form epithelial organoids were compared between PSF and VT endometrial biopsies in a subset of samples. The histology and TEM studies demonstrated relatively mild cellular and sub-cellular damage in both cryopreservation protocols which did not affect tissue functionality and the formation of the organoids. Additionally, the cryopreservation methodology did not affect the gene expression profile of the 68 endometrial-receptivity associated genes studied. In conclusion, our findings indicate that although current cryopreservation methodologies need further improvements, they still allow us to achieve acceptable cell viability and functionality, showing promising potential for facilitating the utilization of cryopreserved endometrial tissue samples for research purposes.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"117 ","pages":"Article 105161"},"PeriodicalIF":2.3,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142567472","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 cryo-facial mask on running performance in amateur middle-distance runners","authors":"Massimo De Nardi ,&nbsp;Luca Filipas ,&nbsp;Simone Di Gennaro ,&nbsp;Silvia Allemano ,&nbsp;Gabriele Gallo ,&nbsp;Andrea Meloni ,&nbsp;Lucio Della Guardia ,&nbsp;Livio Luzi ,&nbsp;Antonio La Torre ,&nbsp;Roberto Codella","doi":"10.1016/j.cryobiol.2024.105158","DOIUrl":"10.1016/j.cryobiol.2024.105158","url":null,"abstract":"<div><div>The excess heat accumulated during exercise can lead to stress-induced fatigue, possibly impairing athletic performance. Various precooling techniques have been applied to enhance thermal comfort, reduce perception of effort, and improve endurance. In this randomized crossover study, twelve male amateur middle-distance runners (age: 33.69 ± 5.9 years; body mass: 71.9 ± 4.4 kg; height: 178.4 ± 5.6 cm; <span><math><mrow><mover><mi>V</mi><mo>˙</mo></mover></mrow></math></span>O₂ peak: 63.3 ± 5.6 mL min⁻¹·kg⁻¹) wore a facial cooling mask before a time-to-exhaustion (TTE) test on a treadmill, under cryostimulation or control conditions. The running performance comprised also two constant load trials, one conducted before and another after wearing the mask, both performed at the velocity of the first ventilatory threshold. Under cryostimulation condition, the TTE was 13 % higher than the control condition (p = 0.0049; <em>d</em> = −0.19) with a significant main effect of time for both ratings of perceived exertion (F_{1, 22} = 50.10; p &lt; 0.0001; η²p = 0.69) and heart rate (F_{1, 22} = 31.53; p &lt; 0.0001; η²p = 0.59). A significant interaction “condition × time” was found for facial skin temperature (F_{2, 44} = 36.93; p &lt; 0.0001; η²p = 0.63) and for heart rate during the constant load trial after wearing the mask (F_{1, 22} = 5.90; p = 0.0238; η²p = 0.21). The localized cryostimulation provided by the mask lowered the skin temperature on the face, potentially mitigating the negative effects of heat stress during running. Incorporating the cryo-facial mask as part of a pre-exercise routine for runners may offer a practical and convenient method to optimize performance and enhance overall training outcomes.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"117 ","pages":"Article 105158"},"PeriodicalIF":2.3,"publicationDate":"2024-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142564154","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":"Vitreous tissue cryopreservation using a blood vessel model and cryomacroscopy for scale-up studies: Observations and mathematical modeling","authors":"Michael J. Taylor ,&nbsp;Prem K. Solanki ,&nbsp;Zhenzhen Chen ,&nbsp;Simona Baicu ,&nbsp;Christina Crossley ,&nbsp;Elizabeth D. Greene ,&nbsp;Lia H. Campbell ,&nbsp;Kelvin G.M. Brockbank ,&nbsp;Yoed Rabin","doi":"10.1016/j.cryobiol.2024.104976","DOIUrl":"10.1016/j.cryobiol.2024.104976","url":null,"abstract":"<div><div>Successful long-term cryobanking of multicellular tissues and organs at deep subzero temperatures calls for the avoidance of ice cryoinjury by reliance upon ice-free cryopreservation techniques. However, the quality of the cryopreserved material is the direct result of its ability to survive a host of harmful mechanisms, chief among which is overcoming the trifecta effects of ice crystallization, toxicity, and mechanical stress. This study aims at exploring improved conditions to scale-up ice-free cryopreservation by combining DP6 as a base cryoprotective agent (CPA) solution with an array of synthetic ice modulators (SIMs). This study is conducted by integrating cryomacroscopy techniques, thermal modeling, solid mechanics analysis, and viability and contractility investigation to correlate physical effects, thermal outcomes, and cryobiology results. As an extension of previous work, this study aims at scale-up of established baseline blood vessel models, while comparing the relative toxicity and vitreous stability of 4 ml and 10 ml samples of DP6 containing either sucrose as a SIM, or the commercial synthetic ice blockers (X1000 and Z1000). Using that established protocol, the addition and removal of DP6+0.6M sucrose and DP6 + 1% X1000 + 1% Z1000 were both well tolerated in rabbit carotid and pig femoral artery models, when assessed for metabolic recovery and contractility. Using cryomacroscopy, it was demonstrated that DP6 + 0.6M sucrose provided a stable vitrification medium under marginal cooling and warming conditions that resulted in &gt;50% survival rate. By contrast, DP6 + 1% X1000 + 1% Z1000 was subject to visible ice formation during cooling under the same thermal conditions, resulting in a significantly lower recovery of ∼20%. Thermal modeling is used in this study to verify the actual cooling and rewarming rates in the specimens, while thermomechanics analysis is used to explain why fractures were observed using cryomacroscopy when the specimens were contained in glass vials but not in plastic vials.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"117 ","pages":"Article 104976"},"PeriodicalIF":2.3,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142371231","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":"Cryopreservation did not affect spermatogonia global methylation profile in Senegalese sole (Solea senegalensis).","authors":"Almeida M Mafalda, Cabrita Elsa, Laizé Vincent, Brionne Aurélien, Labbé Catherine, Fatsini Elvira","doi":"10.1016/j.cryobiol.2024.105162","DOIUrl":"https://doi.org/10.1016/j.cryobiol.2024.105162","url":null,"abstract":"<p><p>Spermatogonia cryopreservation is a method to preserve valuable genomes from both maternal and paternal origin. The damage associated with the application of this technology on post-thaw cell quality is important to assess, including at the epigenetic level. This study aimed to assess post-thawed spermatogonia quality by evaluating alterations in plasma membrane integrity, DNA integrity (fragmentation and apoptosis), lipid peroxidation (malondialdehyde levels) and epigenetic modifications (DNA methylation profile). We observed that plasma membrane integrity (fresh 78.98% ± 5.66; cryopreserved 62.81% ± 3.25; P = 0.003) and DNA integrity (fresh 32.95% ± 2.28; cryopreserved 37.28% ± 1.87; P = 0.0026) were affected by cryopreservation, while no difference in lipid peroxidation was observed (fresh 1.13% ± 0.45; cryopreserved 0.91% ± 0.96; P = 0.701). While global levels of DNA methylation were unaffected by cryopreservation (fresh 82.80% ± 0.47; cryopreserved 83.32% ± 0.81; P = 0.745), some differentially methylated cytosines (DMC) were observed in cryopreserved versus fresh spermatogonia (156 DMC). This study showed that spermatogonia cryopreserved according to our protocol provides a good supply of undamaged cells for several applications. The significance of the few detected DMCs deserves further attention since it may affect gamete differentiation and epigenetic profile.</p>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":" ","pages":"105162"},"PeriodicalIF":2.3,"publicationDate":"2024-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142616413","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":"Cryobiological aspects of upscaling cryopreservation for encapsulated liver cell therapies","authors":"Tom Brookshaw ,&nbsp;Barry Fuller ,&nbsp;Eloy Erro ,&nbsp;Tamnia Islam ,&nbsp;Sweta Chandel ,&nbsp;Elizaveta Zotova ,&nbsp;Clare Selden","doi":"10.1016/j.cryobiol.2024.105155","DOIUrl":"10.1016/j.cryobiol.2024.105155","url":null,"abstract":"<div><div>For the efficient delivery of a cell therapy a treatment must be provided rapidly, at clinical scale, contain a sufficient active cellular component (biomass), and adhere to a multitude of regulatory requirements. Cryopreservation permits many of these demands to be met more readily. Here we present the cryopreservation and recovery of large volume (2.5L) alginate encapsulated liver cell spheroids (AELS), suitable for use with a novel bioartificial liver device (HepatiCan™) for the treatment of those suffering from acute liver failure (ALF), in regulatory approved cryobags and a cryopreservation process optimised for large volumes. By first assessing the thermal profiles of large scale cryobags with a thermal mimic, the feasibility of cryopreserving a full patient dose simultaneously (3x cryobags containing 833 ml biomass each) was investigated, allowing for small and subsequently large-scale testing of cellular functional recoveries. Work presented here demonstrates that optimised reproducible cooling and warming profiles could be achieved with these large volumes, leading to high biomass recoveries at full clinical scale. The recovered AELS also had high regeneration potential, achieving full pre-freeze viable cell densities within 3 days, indicating that the cell therapy could be delivered rapidly to patients with ALF. This study has presented the feasibility for rapid delivery of large volume cell therapies, whilst further research into improved speed of post-thaw recovery is warranted.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"117 ","pages":"Article 105155"},"PeriodicalIF":2.3,"publicationDate":"2024-11-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142496674","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}