Cryobiology最新文献

{"title":"The antioxidant capacity and protective ability of astaxanthin in cryopreservation of mouse spermatogonial stem cells","authors":"Shiva Fathi ,&nbsp;Hassan Nazari ,&nbsp;Mehran Arabi ,&nbsp;Azita Afzali ,&nbsp;Ebrahim Ahmadi","doi":"10.1016/j.cryobiol.2025.105261","DOIUrl":"10.1016/j.cryobiol.2025.105261","url":null,"abstract":"<div><div>Cryopreservation of spermatogonial stem cells (SSCs) offers several benefits, but it can also cause various forms of damage that may reduce the functionality of these cells. Incorporating antioxidants into the cryopreservation medium can provide protection against the detrimental effects of cryopreservation by reducing levels of reactive oxygen species (ROS). In this study, the protective effect of astaxanthin (AST) was evaluated to establish an optimal cryopreservation method for SSCs obtained from the testes of neonatal male mice. AST was added to the freezing base medium at 1, 10, and 100 μM concentrations, and then compared with the control (freezing medium without any additives) and 100 μM vitamin E as a conventional antioxidant. Viability, oxidative stress status, intracellular ROS generation levels, and the expression of <em>Bax</em> and <em>Bcl2</em> were measured in frozen-thawed SSCs 3 weeks after culture and purification. The data showed that the presence of antioxidants, especially 10 μM AST, in the freezing medium significantly increases the viability and total antioxidant capacity (TAC) (P &lt; 0.05), and reduces the levels of lipid peroxidation and intracellular ROS accumulation in the frozen–thawed SSCs. Vitamin E, as well as 10 and 100 μM AST reduced apoptosis in mouse SSCs by downregulating <em>Bax</em> and upregulating <em>Bcl2</em>. The results of this study suggest that adding 10 μM of AST to the freezing medium provides protection to SSCs after thawing. Therefore, the inclusion of 10 μM AST in the cryopreservation medium significantly improves the post-thaw viability and antioxidant capacity of SSCs. These findings indicate that AST could serve as a valuable additive for improving the cryopreservation process of SSCs, thereby offering potential benefits for the preservation of male fertility.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"120 ","pages":"Article 105261"},"PeriodicalIF":2.3,"publicationDate":"2025-06-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144220881","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":"Protein carbonylation reveals the molecular basis of cryoinjury in buffalo spermatozoa","authors":"S.K. Bhure ,&nbsp;D.S. Kumara Wodeyar ,&nbsp;Atul Kumar Sharma ,&nbsp;Abhishek Kumar ,&nbsp;Ajay Kumar ,&nbsp;Manish Mahawar ,&nbsp;S.K. Ghosh ,&nbsp;G. Taru Sharma","doi":"10.1016/j.cryobiol.2025.105271","DOIUrl":"10.1016/j.cryobiol.2025.105271","url":null,"abstract":"<div><div>During cryopreservation, oxidative damage to sperm biomolecules occurs, ultimately compromising its fertilizing ability. Simple proteome analysis may not accurately reflect the cellular functions that are affected, as most carbonylated proteins lose their functions. The carbonylated proteins act as a good marker of oxidative stress. Choosing buffalo spermatozoa as a model, carbonylated proteins from fresh-extended (FESCL) and frozen-thawed (FTSCL) spermatozoa were enriched by affinity chromatography using biotin hydrazide and a monomeric avidin agarose matrix followed by mass spectrometry. Data were analyzed with Proteome Discoverer (v2.2), post-translational modification analysis was performed using SEQUEST, and affected functional activities were predicted using the FunRich tool (v3.0). The mass spectrometric analysis of sperm proteins led to the identification of 415 carbonylated proteins, 151 in FESCL and 405 in FTSCL. Twelve highly abundant carbonylated proteins identified only in FTSCL can be the major contributors to the cryoinjury. The cryopreservation induces the carbonylation of selective sperm proteins. A total of 98 peptides have been precisely annotated that have oxidative modifications. In comparison to FESCL, more proteins in various cellular components, pathways, and processes in FTSCL were carbonylated. The findings indicate the molecular injury to spermatozoa through carbonylated proteins affecting energy metabolism, free-radical scavenging, cytoskeleton, plasma membrane, capacitation, zona-pellucida binding, and sperm-oocyte interaction and are well correlated with the functional parameters of the spermatozoa. The findings provide strong evidence that protein carbonylation is one of the major factors and molecular basis of cryodamage, which compromises the functions of FTSCL. With cryopreservation becoming an inevitable tool, the study becomes more relevant, and this experimental design would give better-quality frozen-thawed spermatozoa with good molecular profiles.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"120 ","pages":"Article 105271"},"PeriodicalIF":2.3,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144205258","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":"Ice ball size and temperature change during cryoablation in a lard and an ethiodized-oil tissue phantom","authors":"Nai-Wen Chang ,&nbsp;Shu-Huei Shen ,&nbsp;Chien-An Liu ,&nbsp;Felix Antony Halim ,&nbsp;Jia-An Hong ,&nbsp;Chih-Chien Li ,&nbsp;Ping-Lang Yen","doi":"10.1016/j.cryobiol.2025.105274","DOIUrl":"10.1016/j.cryobiol.2025.105274","url":null,"abstract":"<div><div>This in vitro study evaluates the effects of varying concentrations of lard and ethiodized-oil (Lipiodol) on temperature changes and ice ball diameters during cryotherapy. A phantom was constructed using six glass bottles, one filled with 0.9 % normal saline (NS) and the others containing agar phantoms mixed with different proportions of lard and NS (0 %, 10 %, 40 %, 70 %, and 100 % lard). Similarly, a phantom was prepared with ethiodized-oil. Six cryoprobes were inserted into the bottles, and freezing was initiated simultaneously, with temperature readings recorded every 10 s over a 9-min freezing period. CT scans were performed before freezing and at 3, 6, and 9 min post-freezing to measure ice ball diameters, with each phantom undergoing a single freezing cycle. Phantoms with lard or ethiodized-oil showed increased freezing rates with higher concentrations, stabilizing at approximately −140 to −150 °C. The largest ice ball diameters were observed in the 0 % and 10 % lard and ethiodized-oil phantoms. Negative correlations were identified between ice ball diameter and lard concentration at 3, 6, and 9 min (R² = 0.910, 0.838, 0.778; p = 0.008, 0.019, 0.030), as well as between ice ball diameter and ethiodized-oil concentration at 6 min (R² = 0.881; p = 0.040). These results suggest that distinct concentrations of lard and ethiodized-oil significantly influence the temporal temperature changes and ice ball dimensions during cryotherapy.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"120 ","pages":"Article 105274"},"PeriodicalIF":2.3,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144205257","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":"Identification of the hub genes involved in ATF5 mediated stress response in vitrified mouse oocytes based on WGCNA","authors":"Guizhen Zhou ,&nbsp;Aiju Liu ,&nbsp;Jiachen Bai ,&nbsp;Yixiao Zhu ,&nbsp;Xinhua Zou ,&nbsp;Yuwen Luo ,&nbsp;Yizaitiguli Reyimjan ,&nbsp;Yan Ma ,&nbsp;Shuaixiang Huang ,&nbsp;Yunpeng Hou ,&nbsp;Jun Li ,&nbsp;Xiangwei Fu","doi":"10.1016/j.cryobiol.2025.105258","DOIUrl":"10.1016/j.cryobiol.2025.105258","url":null,"abstract":"<div><div>Activating transcription factor 5 (ATF5) is a key regulator in the stress response and plays a crucial role in cellular adaptation to the nonphysical environment. Our previous study indicated that a homeostatic state of ATF5 level could improve mitochondrial function in vitrified oocytes. However, the molecular mechanisms underlying the ATF5-mediated gene network in response to oocyte vitrification remain largely unknown. In this study, specific ATF5 siRNA was microinjected into oocytes to suppress ATF5 expression, and oocytes with ATF5 deficiency under normal and vitrification stress conditions were collected for Smart RNA-seq analysis. Weighted gene co-expression network analysis (WGCNA) was used to characterize oocyte co-expression modules involved in the ATF5 mediated response. Our results identified three key gene modules related to ATF5 knockdownRed, Green and, Yellow-using the screening criteria of |R| ≥0.5 and <em>P</em> ≤ 0.05 with WGCNA. Functional enrichment analysis of key gene modules showed that genes in the modules were mainly enriched in apoptosis, ubiquitin mediated proteolysis, the PI3K-Akt signaling pathway, and protein digestion and absorption. STEM and KEGG functional enrichment analysis showed that most genes were involved in protein digestion and absorption, the AMPK signaling pathway, the JAK-STAT signaling pathway, and the apoptosis signaling pathway. Importantly, <em>Diablo</em>, <em>Map3k1</em>, <em>Spta1,</em> and <em>Ubqln4</em>, obtained by sequential scanning of WGCNA and combined functional enrichment analysis, were identified as candidate genes under ATF5 regulation in response to vitrification. These findings demonstrate that the ATF5 mediated gene network exerts regulatory roles in various cellular events and provide novel insights for a deeper understanding of stress response associated impairments in vitrified oocytes.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105258"},"PeriodicalIF":2.3,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144107214","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":"Studies on ovarian tissues’ cryopreservation in the cyprinid species","authors":"Jingting Yao ,&nbsp;Linhui Zeng ,&nbsp;Zheng Zhu ,&nbsp;Ke Feng ,&nbsp;Chaowei Zhou ,&nbsp;Wanliang Wang ,&nbsp;Jianshe Zhou ,&nbsp;Shengqi Su ,&nbsp;Hongyan Xu","doi":"10.1016/j.cryobiol.2025.105251","DOIUrl":"10.1016/j.cryobiol.2025.105251","url":null,"abstract":"<div><div>This study was aimed to establish an effective protocol to cryopreserve the ovarian tissues of cyprinid species, including the crucian carp (<em>Carassius auratus</em>), zebrafish (<em>Danio rerio</em>) and Ya fish (<em>Schizothorax prenanti</em>), crucian carp was used to establish an efficient method of ovarian cryopreservation, zebrafish and Ya fish were used to validate this method. In addition, oxidative stress-related injuries induced by cryopreservation was determined in Ya fish, and the cryopreservation method was modified by adding tea polyphenol (TP) into the cryomedium. First, based on the cellular plasma membrane integrity analysis of the frozen-thawed ovarian tissues, the cryomedium and dehydration/rehydration procedures were intensively evaluated and optimized in the crucian carp. The result showed that the highest cell membrane integrity of cryopreserved ovaries was up to 51.67 ± 0.78 % (including somatic cells and oogonia) after a 7-day cryopreservation under the optimized protocol (the cryomedium containing 10 % FBS, 10 % DMSO, 6 % sucrose, and 6 % BSA in 1xPBS), together with gradient dehydration and rehydration procedures. This protocol was applied to cryopreserve ovaries in another two cyprinid species: in Ya fish<em>,</em> cell membrane integrity of frozen-thawed ovaries was ∼65.12 %, and in zebrafish, it was ∼44.56 %. Moreover, the frozen-thawed ovaries of both two fish species could be used for <em>in vitro</em> culture. Secondly, it was found that the cryopreservation significantly caused oxidative stress (increasing the levels of oxidation index MDA and antioxidant enzymes SOD and CAT) in the frozen-thawed ovaries of Ya fish. Intriguingly, an antioxidant, the TP supplemented into the cryomedium could prominently improve the cells' membrane integrity of frozen-thawed ovaries to 73.82 ± 4.89 %. Likewise, tea polyphenol could also enhance cells’ proliferation potency of the cryopreserved ovaries in Ya fish. In conclusion, in this study, an efficient method was established for cryopreserving fish ovaries, and the findings provided a valuable basis for developing new biological techniques to preserve and exploit the high-quality genetic resources of aquatic species.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105251"},"PeriodicalIF":2.3,"publicationDate":"2025-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144069352","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":"Automated device for rapid sample cooling via controlled submersion","authors":"Purva Joshi ,&nbsp;Zachary Chau ,&nbsp;Shaun Keating ,&nbsp;Colby Langer ,&nbsp;Grace Matheson ,&nbsp;Emily Devorsetz ,&nbsp;Natalie Scanlon ,&nbsp;Mehmet Toner ,&nbsp;Rebecca D. Sandlin","doi":"10.1016/j.cryobiol.2025.105250","DOIUrl":"10.1016/j.cryobiol.2025.105250","url":null,"abstract":"<div><div>An automated device is described which enables programmable submersion in liquid nitrogen to enable rapid specimen cooling for vitrification applications. The device presented here is low-cost, portable, and compatible with a range of cryogenic containers. The device is capable of submerging samples at a range of speeds, enabling the user to optimize the cooling rates based on the thermal mass of the sample as well as the thermal properties of the container and biospecimen. The device consists of a stepper motor that drives a linear actuator, which enables the movement of a 3D-printed robotic arm in the vertical plane which is used to submerge the specimen rapidly into the cryogen. After development, the device was validated for its design parameters. The relative error in starting height and submersion distance was less than 1.5%, indicating a high degree of precision and consistency in positioning during operation. The resulting cooling rates showed no significant difference between manual and automated submersion, confirming the device's performance and reliability. The device performance was further assessed using a 0.25 mL insemination straw to evaluate its practical application. The cooling rate achieved was well within the range cited in previous reports as well as that predicted computationally, confirming the device's functionality. Importantly, this device can be constructed using commercially available materials at relatively low costs.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105250"},"PeriodicalIF":2.3,"publicationDate":"2025-05-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143931833","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":"Levels of post-translationally modified histones in ground squirrel livers are altered during deep torpor","authors":"Remy Carter,&nbsp;Kenneth B. Storey","doi":"10.1016/j.cryobiol.2025.105256","DOIUrl":"10.1016/j.cryobiol.2025.105256","url":null,"abstract":"<div><div>Thirteen-lined ground squirrels (<em>Ictidomys tridecemlineatus</em>) are obligate hibernators capable of reducing their metabolic rates by up to 99 % during winter. Their ability to remain dormant without food for an extended period in cold conditions has made them compelling subjects for research. Developing a clearer understanding of mechanisms surrounding the pre-transcriptional control of hibernating tissues is crucial for cryobiological applications such as organ preservation. Thus, we investigated the differential expression of 24 modified histones (MH) in the livers of torpid and euthermic free-ranging ground squirrels by immunoblotting histone-enriched extracts (p &lt; 0.05). We identified the torpor-responsive downregulation of multiple permissive MHs (H2BK5ac, H3K18ac, H3K23ac, H3K27ac, H3K4me2, H3K4me3, H4K20me1, H4R3me2s), including total H2B and H4, while the linker histone H1.0 was the only histone species that was upregulated. The present study provides valuable insights into the involvement of histone post-translational modifications in the epigenetic landscape of deeply torpid ground squirrel livers.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105256"},"PeriodicalIF":2.3,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143918012","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":"55 MHz constant field dielectric warming of kidneys and ovaries cryopreserved by vitrification","authors":"Brian Wowk ,&nbsp;Alison Ting ,&nbsp;John Phan ,&nbsp;Roberto Pagotan ,&nbsp;Adnan Sharif ,&nbsp;Limdo Chow ,&nbsp;Gregory M. Fahy","doi":"10.1016/j.cryobiol.2025.105257","DOIUrl":"10.1016/j.cryobiol.2025.105257","url":null,"abstract":"<div><div>Organs cryopreserved by vitrification benefit from fast warming to avoid growth and recrystallization of ice that may nucleate during cooling and during warming. Rapid warming is especially important for tissue that doesn't absorb the full concentration of perfused cryoprotectants. Nanowarming uses an oscillating magnetic field to heat magnetic nanoparticles introduced into blood vessels during cryoprotectant perfusion. Dielectric warming uses an oscillating electric field to directly heat water and cryoprotectant molecules everywhere inside an organ. The efficiency of dielectric warming peaks at a particular solution viscosity and temperature that depends on the field oscillation frequency. Below that temperature, field uniformity is very important for uniform warming. An 800 W 55 MHz dielectric warming system was constructed that reached peak warming efficiency at −60 °C instead of −70 °C previously observed at 27 MHz when using the M22 vitrification solution. The shape of capacitor plates that formed the electric field for organ warming was optimized by computer simulation. Computer simulation also provided insights into the effects of organ container shape on internal field uniformity, confirming the theoretical prediction that ellipsoidal shapes are optimum. Dielectric materials surrounding the organ container during warming were used with beneficial effect. In physical experiments with constant field warming at 55 MHz, warming rates peaked near 200 °C/min for ∼15 g rabbit kidneys in 45 mL total volume, and near 700 °C/min for ∼5 g porcine ovaries in 15 mL total volume. Of three rabbit kidneys vitrified, dielectrically warmed under slightly varying conditions, and then transplanted, one survived long-term with return to normal clinical function (serum creatinine &lt;2 mg/dL) in the recipient animal still living 17 months later. The mass of the kidney was 13.9 g, by an order of magnitude the largest vitrified vital organ successfully returned to clinically normal function to date.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105257"},"PeriodicalIF":2.3,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143923659","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 survival after cryopreservation of the endangered exceptional species Crotalaria avonensis","authors":"Megan Philpott ,&nbsp;Anne-Catherine Vanhove ,&nbsp;Mary Chaiken ,&nbsp;Valerie C. Pence","doi":"10.1016/j.cryobiol.2025.105249","DOIUrl":"10.1016/j.cryobiol.2025.105249","url":null,"abstract":"<div><div>Cryopreservation has been shown to be a viable alternative method of long-term storage for exceptional species which cannot be seed banked. While its use is gaining in popularity worldwide for plant conservation, its novelty translates into a dearth of literature on the long-term (&gt;5 year) survival of species in cryo-storage. This study presents an assessment of survival across three variations on the encapsulation vitrification method for 3–16 years of cryostorage for shoot tips of the federally endangered exceptional plant species, <em>Crotalaria avonensis</em>. In addition, the more recently-developed droplet vitrification method was compared to encapsulation vitrification for short-term survival. Overall, over 92 % of banked genotypes survived after long-term cryo-storage, and the droplet vitrification method appears to improve survival over the original encapsulation vitrification procedures. In addition, the long-term survival data validate the previously published guidance that a 40 % initial survival after liquid nitrogen exposure will lead to a 95 % chance of survival after long-term storage, with 39 % initial survival in <em>C. avonensis</em> leading to a 95 % probability of survival after long-term storage per genotype. Initial survival after short-term liquid nitrogen exposure appears to be an indicator of long-term survival in storage, which can give practitioners some indication of their banking success. This study contributes to the literature showing the potential and success of using cryopreservation as a long-term banking method for threatened and/or wild exceptional species.</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105249"},"PeriodicalIF":2.3,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143918009","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":"Morphological characteristics, mitochondrial oxidoreductase activity, and vascularization of human adipose tissues pre-and post-xenotransplantation into BALB/c female nude mice","authors":"Banafsheh Heidari ,&nbsp;Negar Eidi ,&nbsp;Pejman Mortazavi ,&nbsp;Zahra Saffarian ,&nbsp;Azadeh Soltani ,&nbsp;Parvin Mansouri ,&nbsp;Seyed Mehdi Tabaie","doi":"10.1016/j.cryobiol.2025.105248","DOIUrl":"10.1016/j.cryobiol.2025.105248","url":null,"abstract":"<div><div>This study aimed to evaluate the effect of different cryoprotectants on the histomorphological characteristics, oil ratio (OR) indices, mitochondrial oxidoreductase activities, and vascularization of human adipose tissues after thawing (pre-xenotransplantation), one month and three months post-xenotransplantation. For this purpose, we froze human adipose tissues in five experimental groups, including NCG (PBS without any cryoprotectants), FG1 (4 % DMSO+6 % Trehalose), FG2 (6 % Trehalose+2 % DMSO+9 % FBS), FG3 (2 % DMSO+4 % Trehalose+3 % Sucrose+9 % FBS), and FG4 (4 % DMSO+3 % Trehalose+3 % Sucrose) and compared them with the positive control group (PCG or fresh adipose tissue). Adipose tissues were frozen with a cooling rate of −1 °C/min and stored in a freezer at −20 °C until use. Three months after freezing, human adipose tissues were xenotransplanted into the BALB/c nude mice for one and three months. Then, xenografts were removed and evaluated for histopathological properties, OR indices, mitochondrial activities, and vascularization. After thawing (pre-xenotransplantation) and post-xenotransplantation (one- and three-month after xenografting), the NCG group demonstrated the most significant tissue damage including necrosis, multiple cysts, inflammatory cell invasion, cell rupture, fibrosis, necrosis and ECM disintegration (<em>p ≤</em> 0.05). Pre-xenotransplantation, one- and three-month after grafting, the best structural integrity of adipose tissues with the least degeneration of adipocytes was detected in the FG2 and FG3 groups (<em>p ≤</em> 0.05). Pre- and post-xenotransplantation, the highest and least OR indices and mitochondrial oxidoreductase activities were demonstrated in the NCG and FG2 groups, respectively (<em>p ≤</em> 0.05). The least VEGF and CD31 expressions were shown in the NCG group at pre- and post-transplantation (<em>p ≤</em> 0.05). The highest CD31 and VEGF expressions were detected in the FG3 and FG2 groups after thawing and grafting (both one- and three-month) (<em>p ≤</em> 0.05).</div></div>","PeriodicalId":10897,"journal":{"name":"Cryobiology","volume":"119 ","pages":"Article 105248"},"PeriodicalIF":2.3,"publicationDate":"2025-05-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143911627","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}