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Background: In the cryopreservation process, rooster spermatozoa are vastly sensitive to reactive oxygen species (ROS). This study aimed to investigate the effects of Lake extender supplemented via Cysteamine (CYS) on the quality and fertility characteristics of rooster semen during the cryopreservation process. Methods: Semen samples were collected from 10 proved Ross-308 roosters, diluted and cryopreserved in the Lake extender which contained 0, 1, 2, 4, and 8 mM of CYS (C-0, C-1, C-2, C-4, and C-8, respectively). Motility parameters, membrane integrity, abnormal morphology, mitochondrial activity, acrosome integrity, viability, apoptosis status, lipid peroxidation, DNA fragmentation, ROS concentration, as well as fertility potential were evaluated after thawing. Results: total motility and progressive motility were higher (P ≤ 0.05) in C-1 and C-2 compared to the other groups. The C-1 showed higher (P ≤ 0.05) membrane integrity, mitochondrial activity, acrosome integrity, viability, and lower (P ≤ 0.05) late apoptosis, lipid peroxidation, DNA fragmentation, and ROS concentration compared to the other groups. During fertility evaluation, C-1 presented a higher fertility rate than the control group. In cases of velocity parameters, abnormal morphology, early apoptosis, necrosis, and hatching rate, no significant difference (P > 0.05) was found. Conclusion: using the optimal concentration of CYS (1 mM) in the Lake extender is useful for protecting rooster sperm during the cryopreservation process and it could be used for reproductive programs. [ABSTRACT FROM AUTHOR]

The present study explores the advantages of enriching the freezing medium with membrane lipids and antioxidants in improving the outcome of prepubertal testicular tissue cryopreservation. For the study, testicular tissue from Swiss albino mice of prepubertal age group (2 weeks) was cryopreserved by slow freezing method either in control freezing medium (CFM; containing DMSO and FBS in DMEM/F12) or test freezing medium (TFM; containing soy lecithin, phosphatidylserine, phosphatidylethanolamine, cholesterol, vitamin C, sodium selenite, DMSO and FBS in DMEM/F12 medium) and stored in liquid nitrogen for at least one week. The tissues were thawed and enzymatically digested to assess viability, DNA damage, and oxidative stress in the testicular cells. The results indicate that TFM significantly mitigated freeze–thaw-induced cell death, DNA damage, and lipid peroxidation compared to tissue cryopreserved in CFM. Further, a decrease in Cyt C, Caspase-3, and an increase in Gpx4 mRNA transcripts were observed in tissues frozen with TFM. Spermatogonial germ cells (SGCs) collected from tissues frozen with TFM exhibited higher cell survival and superior DNA integrity compared to those frozen in CFM. Proteomic analysis revealed that SGCs experienced a lower degree of freeze–thaw-induced damage when cryopreserved in TFM, as evident from an increase in the level of proteins involved in mitigating the heat stress response, transcriptional and translational machinery. These results emphasize the beneficial role of membrane lipids and antioxidants in enhancing the cryosurvival of prepubertal testicular tissue offering a significant stride towards improving the clinical outcome of prepubertal testicular tissue cryopreservation. [ABSTRACT FROM AUTHOR]

Liver biobanking is a promising approach that saves the lives of patients with end-stage liver disease. Cryopreservation based on vitrification enables semi-permanent organ preservation, contributing to overcome the shortage of donors for liver transplants. A technical challenge in cryopreservation of transplantable organs lies in thawing methodology, and conventional convective warming cannot maintain the glassy state during thawing because of the large temperature gradient between the inner and outer parts of the organs, leading to ice formation and damage of cells in the organ. Nanowarming, in which magnetic nanoparticles are dispersed in a vitrification solution and heated by exposure of alternating magnetic field, can achieve uniform and rapid heating of organs. Herein, we report that amphiphilic phospholipid polymers composed of 2-methacryloyloxyethyl phosphorylcholine and n -butyl methacrylate can function as a cryoprotectant for nanowarming. The amphiphilic phospholipid polymers enhanced the viability of primary rat hepatocytes after vitrification. Moreover, the polymers enhanced the dispersion stability of magnetic nanoparticles in vitrification solution, and the perfusion of the vitrification solution with magnetic nanoparticles into rat livers through portal vein provided uniform distribution of the nanoparticles in the liver. After perfusion, the vitrified liver was successfully thawed rapidly and uniformly by nanowarming, which maintained tissue integrity and cell viability. [ABSTRACT FROM AUTHOR]

Prunella vulgaris L.(PV) and Tussilago farfara (TF) are perennial herbs rich in flavonoids and phenolic compounds with immense medicinal value. PV extract (PV-E) possesses potent antipyretic, anti-inflammtory, antioxidant, antiseptic, anti-cancer and immune stimulatory properties and have been traditionally known for the treatment of wounds, ulcers and sores. TF extract (TF-E) has been known for antibacterial, antioxidant, anti-inflammatory, anti-viral, anti-diabetic, anti-cancer, anti-obesity and wound healing effects. Additionally, TF-E infusions have been used for asthma, cough, and bronchopneumonia treatments. The therapeutic efficacy of transplanted human adipose stem cells (hASCs) is abrogated under the deteriorating effects of heat stress offered by burn wounds. Earlier researches has documented antioxidant priming as an effective strategy to enhance stem cell performance. As both PV-E and TF-E are known for their potent antioxidant effects. The present study aims to examine the cryoprotective effects of PV-E and TF-E priming on hASCs against in-vitro heat-induced thermal stress. Moreover, we determined the anti-inflammatory potential of both PV-E and TF-E on rabbits. Antioxidant capacity of both PV-E and TF-E is examined via DPPH assay and anti-inflammatory activity is assessed in rabbits using carrageen-induced paw edema model of inflammation. Next, we investigate the efficacy of different doses (1.25–100 μg/ml) of PV-E and TF-E on hASCs; MTT, LDH, calcein AM staining, and wound scratch assay were used to assess cell viability, cytotoxicity, proliferation ability and cell migration potential in the cells. Then, hASCs were pretreated for 24 h with optimum doses of PV-E and TF-E determined from MTT assay results and were subsequently exposed to in-vitro thermal injury (51 °C,10 min). The cytoprotective effects of both PV-E and TF-E priming under thermal stress were investigated via MTT, LDH, annexin-V staining and gene expression analysis. Both PV-E and TF-E extracts demonstrated potent antioxidant and effective anti-inflammatory activities, with a clear reduction in inflammation. Study on hASCs exhibited improved cell viabilities, enhanced cell proliferation and migration abilities of both extracts. While heat stress data revealed that PV-E (2.5 μg/ml) and TF-E (5 μg/ml) pretreatment significantly ameliorated effects of thermal-injuries in hASCs as depicted by significantly enhanced cell viabilities, low LDH release profile, and lower annexin-V expression and regulated gene expression of the pretreated cells. PV-E and TF-E priming effectively enabled hASCs to combat thermal injury by significantly promoting cell survival than untreated cells. Hence, these findings suggest that PV-E and TF-E priming could be used to attain improved cellular responses and enhanced therapeutic efficacy in burnt tissue. [Display omitted] • Methanolic extracts of Prunella vulgaris (PV-E) and Tussilago farfara (PV-E) possess potent antioxidant and anti-inflammatory effects. • PV-E and TF-E significantly enhanced proliferation and migration potential of hADSs. • HASCs primed with PV-E and TF-E effectively combat thermal injury as compared to untreated hASCs. • PV-E and TF-E can serve as a priming agent in stem cell therapies to improve their survival and regenerative capabilities. [ABSTRACT FROM AUTHOR]

Protein deterioration caused by ice crystals is an important factors affecting the frozen storage of fish. In this study, antifreeze peptides extracted from hydrolysates of sea cucumber intestinal protein with inhibition of protein denaturation were screened and characterized. The peptide Leu-Pro-Glu-Phe-Thr-Glu-Glu-Glu-Lys (LPEFTEEEK), derived from neutral protease hydrolysates of sea cucumber intestinal protein, was investigated for its potential to enhance the quality of salmon fillets during three freeze-thaw cycles. The results showed that the application of LPEFTEEEK effectively maintained the texture of fish fillets, as well as the oxidative and conformation stability of myofibrillar protein during the freezing process. Additionally, molecular dynamics simulations verified that LPEFTEEEK could bind to ice crystals and inhibit their recrystallization, thus preventing organisms from being damaged by freezing. This suggests that LPEFTEEEK holds significant promise as a novel cryoprotective agent for marine-derived antifreeze peptides. [Display omitted] • A novel antifreeze peptide LPEFTEEEK was screened from sea cucumber intestinal. • Peptide LPEFTEEEK can maintain the quality of fish during the freeze-thaw cycle. • Peptide LPEFTEEEK can inhibit ice crystal growth through interaction. • Peptide LPEFTEEEK can reduce the destruction of protein structure by ice crystals. [ABSTRACT FROM AUTHOR]

Cryopreservation facilitates long-term semen storage and enables the exchange of genetic material among elite livestock over extensive distances. A decrease in sperm quality is an unavoidable outcome of the cryopreservation process. Prior research has established that incorporating cryoprotectants into the diluent can mitigate freeze-induced damage and enhance sperm quality. This study aims to assess the impact of Epimedium polysaccharide (EPS) on the cryopreservation of goat semen. Samples were obtained from six healthy goats following an initial examination. One portion of the semen was diluted with a base solution containing EPS for treatment purposes, whereas another was diluted without EPS, serving as the control. Results indicated that varying concentrations (1, 2, 3, 4, 5 mg/mL) of EPS in the diluent enhanced both physiological characteristics and antioxidant enzyme activities in cryopreserved goat sperm. Further analysis showed that the 3 mg/mL EPS concentration significantly improved sperm motility (52.10 %), plasma membrane integrity (57.01 %), tail plasma membrane integrity (52.37 %), acrosome integrity rate (52.45 %), and antioxidant enzyme activities relative to other groups (P < 0.05). Additionally, the inclusion of 3 mg/mL EPS substantially improved the sperm's fertilization capability. In conclusion, our experiments confirm that EPS supplementation significantly enhances sperm quality post-freezing, with 3 mg/mL identified as the optimal concentration. [ABSTRACT FROM AUTHOR]