Aseptic capillary vitrification of human spermatozoa: cryoprotectant-free vs. cryoprotectant-included technologies.
Abstract The protocol of aseptic cryoprotectant-free vitrification on human spermatozoa is well documented. However, data about the effect of permeable cryoprotectants at this procedure is limited. Presented study aimed to test the aseptic capillary vitrification technologies using permeable cryoprotectant-included or cryoprotectant-free media. Thirty-two normal samples were included and analyzed after vitrification in three different media and thawing. Three treatment groups were formed: Group 1, basic medium; Group 2, basic medium with 0.25 M sucrose; Group 3, basic medium with glycerol. Before plunging into liq...
Source: Cryobiology - January 7, 2021 Category: Biology Authors: Wang M, Todorov P, Isachenko E, Rahimi G, Wang W, von Brandenstein M, Kumar P, Mallmann P, Isachenko V Tags: Cryobiology Source Type: research

Tumor location on MRI determines outcomes of patients with prostate cancer after total prostate cryoablation.
Abstract We investigated the association between tumor location on multiparametric magnetic resonance imaging (mpMRI) and outcomes of prostate cancer patients after primary total prostate cryoablation (PTPC). Between March 2010 and December 2012, consecutive 192 prostate cancer patients receiving PTPC were enrolled. Tumor locations were determined and classified as anterior apex (AA), anterior midgland (AM), anterior base (AB), posterior apex (PA), posterior midgland (PM) and posterior base (PB) using mpMRI. Midline location, central location, seminal vesicle invasion, extraprostatic extension, multiple tumors, an...
Source: Cryobiology - January 5, 2021 Category: Biology Authors: Chen CH, Chen YC, Pu YS Tags: Cryobiology Source Type: research

Resistance to ventricular fibrillation predicted by the QRS/QTc - ratio in an intact rat model of hypothermia/rewarming.
Abstract Accidental hypothermia is associated with increased risk for arrhythmias. QRS/QTc is proposed as an ECG-marker, where decreasing values predict hypothermia-induced ventricular arrhythmias. If reliable it should also predict nonappearance of arrhythmias, observed in species like rat that regularly tolerate prolonged hypothermia. A rat model designed for studying cardiovascular function during cooling, hypothermia and subsequent rewarming was chosen due to species-dependent resistance to ventricular arrhythmias. ECG was recorded throughout the protocol. No ventricular arrhythmias occurred during experiments...
Source: Cryobiology - January 4, 2021 Category: Biology Authors: Dietrichs ES, Selli AL, Kondratiev T, McGlynn K, Smith G, Tveita T Tags: Cryobiology Source Type: research

Effect of exogenous lipids on cryotolerance of Atlantic salmon (Salmo salar) spermatozoa.
This study investigated the effect of exogenous lipids in the freezing medium on the post-thaw functional parameters of Salmo salar spermatozoa. Semen samples (n = 12) were incubated in standard extender supplemented with different concentrations of oleic acid (OA, C18:1n9), linoleic acid (LA, C18:2n6), arachidonic acid (ARA, C20:4n6) and cholesterol-loaded cyclodextrin (CLC). Post-thaw motility, membrane integrity, mitochondrial membrane potential (ΔΨm), superoxide anion (O2•-) and fertility rates were analysed. The results revealed that the semen incubated with 0.003 mmol/L OA increased the motility (∼...
Source: Cryobiology - January 4, 2021 Category: Biology Authors: Díaz R, Quiñones J, Short S, Contreras P, Ulloa-Rodríguez P, Cancino-Baier D, Sepúlveda N, Valdebenito I, Farías JG Tags: Cryobiology Source Type: research

Thermomechanical stress analysis of rabbit kidney and human kidney during cryopreservation by vitrification with the application of radiofrequency heating.
This study presents a computational framework for thermomechanical stress analysis in a specimen undergoing cryopreservation, with emphasis on radiofrequency (RF) heating for recovering from cryogenic storage. In particular, this study addresses cryopreservation by vitrification, where the specimen is stored in the amorphous phase (vitreous means glassy). In broad terms, the relatively high cooling and rewarming rates necessary for vitrification result in differential thermal expansion in the specimen, which is the driving force for thermomechanical stress. Thermomechanical stress can lead to structural damage, such as fra...
Source: Cryobiology - January 4, 2021 Category: Biology Authors: Solanki PK, Rabin Y Tags: Cryobiology Source Type: research

Cryopreservation of small numbers of human spermatozoa in a Stripper tip: report of the first live-birth worldwide.
CONCLUSIONS: This is the first successful delivery following the use of spermatozoa frozen in an original device, the Stripper Tip, giving a promising prospect for managing severe male infertilities. PMID: 33400960 [PubMed - as supplied by publisher] (Source: Cryobiology)
Source: Cryobiology - January 2, 2021 Category: Biology Authors: Herbemont C, Mnallah S, Bennani-Smires B, Peigne M, Cedrin-Durnerin I, Grynberg M, Sifer C Tags: Cryobiology Source Type: research

Effect of organic selenium dietary supplementation on quality and fertility of cryopreserved chicken sperm.
Abstract Oxidative stress due to cryopreservation has been considered as a major factor in sperm damage. Supplementation of the diet with different concentrations of organic selenium has been proposed to improve the quality of fresh and frozen-thawed semen in different breeds of roosters. Sixteen Pradu Hang Dum (Thai native) and 16 Rhode Island Red roosters were used in this study. Four levels of selenium supplementation between 0 and 0.9 ppm were examined. After 14 days of feeding, semen samples were collected twice a week and the fresh semen was evaluated. Then semen from each group was pooled and cryopreserved....
Source: Cryobiology - January 2, 2021 Category: Biology Authors: Chauychu-Noo N, Thananurak P, Boonkum W, Vongpralub T, Chankitisakul V Tags: Cryobiology Source Type: research

Enhancing the preservation of liposomes: the role of cryoprotectants, lipid formulations and freezing approaches.
Abstract In the last decades, liposomes acquired a striking success in the biomedical field thanks to their biocompatibility and drug delivery ability. Many liposomal drug formulations have been already approved by the Food and Drug Administration (FDA) and used for the treatment of a wide range of pathologies with or without further engineering. Their clinical application requires strict compliance with high standard quality rules, and it is crucial to employ storage methods that do not affect the integrity of the vesicles and preventing the leakage of their cargo. In this work, the design of a suitable formulati...
Source: Cryobiology - January 2, 2021 Category: Biology Authors: Susa F, Bucca G, Limongi T, Cauda V, Pisano R Tags: Cryobiology Source Type: research

Supplementation of exogenous lipids via liposomes improves coral larvae settlement post-cryopreservation and nano-laser warming.
Abstract Coral reefs worldwide are receding because of detrimental human activities, and cryopreservation of coral larvae would ensure that their genetic biodiversity is not irremediably lost. In recent years, the vitrification and laser warming of coral propagules has demonstrated promising results. During cryopreservation, cellular membranes undergo substantial reconfigurations that may affect survival. Fat enrichment may alter the physical proprieties of cell membranes and improve resistance to low temperatures. Therefore, the aim of this study was to determine whether supplementation of exogenous lipids using ...
Source: Cryobiology - December 29, 2020 Category: Biology Authors: Cirino L, Tsai S, Wang LH, Chen CS, Hsieh WC, Huang CL, Wen ZH, Lin C Tags: Cryobiology Source Type: research

Establishment, characterization, and cryopreservation of cell lines derived from red-rumped agouti (Dasyprocta leporina Linnaeus, 1758) - A study in a wild rodent.
In conclusion, we have shown that viable fibroblasts can be obtained from red-rumped agouti skin, featuring minimal changes after eight passages in in vitro culture systems. Additionally, adjustments to the cryopreservation protocol are necessary to reduce cellular oxidative stress caused by low temperatures. PMID: 33359644 [PubMed - as supplied by publisher] (Source: Cryobiology)
Source: Cryobiology - December 24, 2020 Category: Biology Authors: Praxedes ÉA, Silva MB, Medeiros de Oliveira LR, Vitor da Silva Viana J, Silva AR, Franco de Oliveira M, Pereira AF Tags: Cryobiology Source Type: research

Sonoporation Enables High-Throughput Loading of Trehalose into Red Blood Cells.
Abstract Despite recent advances in biostabilization, clinical blood supplies still experience shortages and storage limitations for red blood cells (RBCs) have not yet been sufficiently addressed. Storing RBCs in a frozen or dried state is an appealing solution to address storage limitations, but many promising cryoprotectants, including the non-reducing sugar trehalose, are impermeant to mammalian cell membranes and cannot be utilized effectively using currently available compound-loading methods. We found that transient pore formation induced by ultrasound and microbubbles (sonoporation) offers an effective mea...
Source: Cryobiology - December 24, 2020 Category: Biology Authors: Janis BR, Priddy MC, Otto MR, Kopechek JA, Menze MA Tags: Cryobiology Source Type: research

ApSerpin-ZX from Agapanthus praecox, is a potential cryoprotective agent to plant cryopreservation.
This study indicated that ApSerpin-ZX was a potential cryoprotective agent that alleviated oxidative stress and cell death induced by cryopreservation. PMID: 33316226 [PubMed - as supplied by publisher] (Source: Cryobiology)
Source: Cryobiology - December 11, 2020 Category: Biology Authors: Chen G, Li R, Shen X Tags: Cryobiology Source Type: research

Effect of cryopreservation on the bacterial community structure of filamentous cyanobacteria, Trichormus variabilis (Nostocales, Cyanobacteria).
In this study, research on the changes in co-cultured bacterial community structure during cyanobacterial cryopreservation were investigated using three different experimental groups by next generation sequencing (NGS): 1) cultured Trichormus variabilis without cryopreservation (control group), 2) cultured T. variabilis after cryopreservation in 10% dimethyl sulfoxide (Me2SO) for 14 days (cryo-cell group), and 3) cultured T. variabilis after cryopreservation in 10% Me2SO for 14 days within alginate beads (cryo-bead group). The results showed that the abundance of Sphingomonas and Hydrogenophaga (belonging to phylum Proteob...
Source: Cryobiology - December 9, 2020 Category: Biology Authors: Park M, Kim M, Park T, Lee CS Tags: Cryobiology Source Type: research

Cryobanking of primary somatic cells of elite farm animals - a pilot study in domesticated water buffalo (Bubalus bubalis).
This study aimed to 1) establish and cryopreserve somatic cells from elite buffaloes, and 2) share stored somatic cells and their associated data with researchers. To achieve these targets, somatic cells were established successfully from tail-skin biopsies of 17 buffaloes. The informative data such as buffalo details (breed, date of birth, sex, and age at the time of tissue biopsy collection, and production traits), the number of cryovials stored, and freezing dates were recorded in an electronic file and a printed inventory record. The established somatic cells were flat, spindle-shaped morphology, and expressed vimentin...
Source: Cryobiology - December 7, 2020 Category: Biology Authors: Dua S, Sharma P, Saini M, Rawat N, Rajendran R, Bansal S, Wakil AM, Beniwal M, Parashar A, Bajwa KK, Selokar NL, Kumar R, Kumar D, Yadav PS Tags: Cryobiology Source Type: research

Molecular cloning, characterization and expression analysis of CbPLD δ gene from Chorispora bungeana in low temperature.
In this study, one member of CbPLD gene family, CbPLDδ, was cloned from C. bungeana and was introduced into tobacco. This gene encodes an 864-amino acid protein with two catalytic HxKxxxxD motifs which are essential for phospholipase D activity. After the CbPLDδ gene is fused with the vector containing the GFP tag, subcellular localization showed that CbPLDδ was predominately located in the cell membrane. RT-qPCR and histochemical GUS assays showed that CbPLDδ gene was induced by low temperature and expressed predominantly in leaf and root. Compared with wild-type tobacco, CbPLDδ transgenic to...
Source: Cryobiology - December 5, 2020 Category: Biology Authors: Yang N, Liu B, Yang P, Chen X, Li H, Wang W, Li Q, Wang S Tags: Cryobiology Source Type: research

Equilibrium vitrification of mouse embryos using low concentrations of cryoprotectants.
In this study, we examined whether embryos could be vitrified in a near-equilibrium state using a solution containing low concentrations of cryoprotectants and thus with low osmolality. To investigate whether embryos were vitrified in a near-equilibrium state, 2-cell mouse embryos were vitrified with EDFS10/10a, PB1 medium containing 10% (v/v) ethylene glycol, 10% (v/v) DMSO, and 0.4 M sucrose, in liquid nitrogen, stored at -80oC for 4-28 days, and warmed in water at 25oC. The viability of the embryos was evaluated by the appearance of embryos after warming and developmental ability. When embryos were vitrified in liquid n...
Source: Cryobiology - December 4, 2020 Category: Biology Authors: Qiu J, Hasegawa A, Mochida K, Ogura A, Koshimoto C, Matsukawa K, Edashige K Tags: Cryobiology Source Type: research

Effect of cryopreservation on pollen viability, fertility and morphology of different Psidium species.
Abstract Cryopreservation of pollen is a complementary conservation strategy and can be used for conserve the diversity in the genus Psidium. The present study aims to cryopreserve the pollen of Psidium species to overcome asynchronous flowering. The pollen of different Psidium species were germinated in vitro in an optimized medium of germination. In vitro/in vivo pollen viability assessment and SEM analysis were carried out to determine the changes after cryopreservation. The in vitro pollen viability was determined at monthly intervals starting from fresh pollen until six months of cryopreservation. The in vivo...
Source: Cryobiology - December 4, 2020 Category: Biology Authors: Vishwakarma PK, Vincent L, Vasugi C, Rajasekharan PE Tags: Cryobiology Source Type: research

Response of Anatolian mountain frogs (Rana macrocnemis and Rana holtzi) to freezing, anoxia, and dehydration: Glucose as a cryoprotectant.
In conclusion, our results suggest that glucose may be identified as an important cryoprotectant that plays an important role in the survival of Anatolian mountain frogs during extreme conditions. PMID: 33285112 [PubMed - as supplied by publisher] (Source: Cryobiology)
Source: Cryobiology - December 4, 2020 Category: Biology Authors: Yoldas T, Cengiz Erismis U Tags: Cryobiology Source Type: research

Physiological responses to cryoprotectant treatment in an early larval stage of the malaria mosquito, Anopheles gambiae.
This study provides ample evidence for an active osmoregulatory response in the Anopheles larvae to counter the permeation of cryoprotectants from the surrounding medium. The data shows a strong correlation between the larval mortality and water content, indicating an osmoregulatory crisis in the larva due to certain cryoprotectants such as the higher concentrations of ethane diol (ED). The observations also indicate that the ability of the larvae to regulate permeation and water balance ceases at or within 20 minutes of cryoprotectant exposure, but this is strongly influenced by the treatment temperature. Among the compou...
Source: Cryobiology - December 3, 2020 Category: Biology Authors: Campbell JB, Dosch A, Hunt CM, Dotson EM, Benedict MQ, Rajamohan A, Rinehart JP Tags: Cryobiology Source Type: research

Melatonin induction of HSP90 expression exerts cryoprotective effect on ovarian tissue.
Abstract Our previous study revealed that melatonin (MLT) protected the quality of cryopreserved ovarian tissues in mice. This work was carried out to examine the role of MLT in inducing HSP90 expression of ovarian tissue for achieving cryoprotection. Pieces of ovarian tissues were obtained from 50 female rats treated with MLT at 0, 0.001, 0.01, 0.1, and 1 mM, respectively. After cryopreservation-thawing, HSP90 mRNA and protein level were evaluated using qRT-PCR and western blot. The qRT-PCR results revealed that HSP90 mRNA expression was significantly (p
Source: Cryobiology - December 3, 2020 Category: Biology Authors: Cheng LY, Sun TC, Liu XC, Yu H, Zhou SJ, Tian L, Yang SH, Liu BX Tags: Cryobiology Source Type: research

Cryopreservation of mesenchymal stromal cell-derived extracellular vesicles using trehalose maintains their ability to expand hematopoietic stem cells in vitro.
Abstract The multitude of clinical trials using mesenchymal stromal cells (MSCs) has underscored their significance as a promising cell source for regenerative therapies. Most studies have however shown that MSCs get entrapped into the microvasculature of lungs, liver and spleen. In addition to intercellular communication, MSCs exert their effects in a paracrine manner by secretion of extracellular vesicles (EVs). The therapeutic effects of MSC-derived EVs have been examined in several diseases such as hepatic failure, liver injury, hematopoiesis etc. Therefore, optimization of cryopreservation strategies for the ...
Source: Cryobiology - November 27, 2020 Category: Biology Authors: Budgude P, Kale V, Vaidya A Tags: Cryobiology Source Type: research

Insights into the model of non-perfect osmometer cells for cryopreservation: a parametric sweep analysis.
In conclusion, cell volume at equilibrium depends on the position of the temporal window of mechanosensitive channels opening, which, in general, is a complex function of model parameters and operating conditions. PMID: 33248930 [PubMed - as supplied by publisher] (Source: Cryobiology)
Source: Cryobiology - November 26, 2020 Category: Biology Authors: Traversari G, Cincotti A Tags: Cryobiology Source Type: research

Maturation and fertilization of African lion (Panthera leo) oocytes after vitrification.
o M Abstract The African lion is an excellent model species for the highly endangered Asiatic lion. African lions reproduce well in zoos, leading to the fact that occasionally ovaries and testis are available for in-vitro experiments. We previously performed in-vitro maturation (IVM) and fertilization of lion oocytes and were able to produce advanced embryos after intracytoplasmic sperm injection (ICSI) with cryopreserved sperm. Here we examined whether our in-vitro method is also applicable after vitrification of immature oocytes. Oocytes of four lionesses (5-7 years old) were obtained after euthanasia and immedi...
Source: Cryobiology - November 25, 2020 Category: Biology Authors: Zahmel J, Jänsch S, Jewgenow K, Sandgreen DM, Simonsen KS, Colombo M Tags: Cryobiology Source Type: research

Antioxidants and their effect on the oxidative/nitrosative stress of frozen-thawed boar sperm.
ute;nchez R Abstract In swine, the use of frozen-thawed boar sperm for artificial insemination remains a suboptimal reproductive technology. Among the negative effects of cryopreservation on sperm cells, it is worth highlighting that cryopreservation causes irreversible alterations in motility and components of the sperm membrane as a result of dramatic changes in temperature (cooling/freezing curve) and osmolality. In addition, freeze-thawing may induce oxidative stress and increase the generation of reactive oxygen species (ROS) and nitrogen reactive species (RNS). While boar sperm cryopreservation has been repo...
Source: Cryobiology - November 25, 2020 Category: Biology Authors: Pezo F F, Yeste M, Zambrano F, Uribe P P, Risopatrón J, Sánchez R Tags: Cryobiology Source Type: research

Major Challenges in Cryopreservation of sea urchin eggs.
Abstract Cryopreservation of gametes, embryos and larvae of marine invertebrates has been investigated in many studies throughout the years. There are many favorable studies on sperm cryopreservation but oocytes are still under research as no successful results have been sustainably obtained for this type of cells. The preservation of both maternal and paternal gametes separately would provide a reliable source of genetic material for their application to conservation, aquaculture and fundamental research. Unfortunately to date, it has not been possible to cryopreserve eggs from marine organisms. The aim of this r...
Source: Cryobiology - November 25, 2020 Category: Biology Authors: Campos S, Troncoso J, Paredes E Tags: Cryobiology Source Type: research

Cryotherapy mediates histopathological and microstructural changes during the treatment of skin and subcutaneous tumors in dogs.
Abstract The therapeutic effects of cryotherapy on skin and subcutaneous tumors in dogs were retrospectively studied in 20 dogs with 37 tumor lesions, of which 30 were benign and seven were malignant. Our results showed that during follow-up, 94.5% of lesions were completely exfoliated, without relapse or metastasis (mean time = 245.7 days). To investigate the effects of cryotherapy, we compared histopathological observations and microstructural changes in healthy tissues and tumor tissues, before and after cryotherapy. After cryotherapy, both normal skin and tumor tissue exhibited edema and hyperemia, with inflam...
Source: Cryobiology - November 25, 2020 Category: Biology Authors: Ma J, Yu X, Lv J, Lin D, Lin J, Bai Y, Wang Y, Li X, Dong J Tags: Cryobiology Source Type: research

Effect of sulforaphane on apoptosis, reactive oxygen species and lipids peroxidation of human sperm during cryopreservation.
Abstract Sperm cryopreservation is a common procedure to preserve viable sperm for an indefinite period. This procedure has numerous detrimental effects on sperm function due to increased generation of reactive oxygen species (ROS). During cryopreservation, while ROS increases, antioxidant enzymes level decreases. It has been shown that a relationship exist between lower antioxidant levels and infertility. L-Sulforaphane (SFN) is an isothiocyanate in cruciferous vegetables of the brassica class that has potent protective effects against oxidative stress. The purpose of the present study was to evaluate the effects...
Source: Cryobiology - November 25, 2020 Category: Biology Authors: Valipour J, Nashtaei MS, Khosravizadeh Z, Mahdavinezhad F, Nekoonam S, Esfandyari S, Amidi F Tags: Cryobiology Source Type: research

Optimization of UC-MSCs cold-chain storage by minimizing temperature fluctuations using an automatic cryopreservation system.
Abstract The effective long-term cryopreservation of human mesenchymal stem cells is an essential prerequisite step and represents a critical approach for their sustained supply in basic research, regenerative medicine, and tissue engineering applications. Off-the-shelf availability of human umbilical cord-derived mesenchymal stromal cells (UC-MSCs) for regenerative medicine application requires the development of nontoxic, safe, and efficient protocols for cryopreservation. In the long-term low-temperature storage process of cells, traditional manual storage has a great impact on cell activity, recovery, and func...
Source: Cryobiology - November 25, 2020 Category: Biology Authors: Xu Y, Li-Ying Chan L, Chen S, Ying B, Zhang T, Liu W, Guo H, Wang J, Xu Z, Zhang X, He X Tags: Cryobiology Source Type: research

Protective effects of Tribulus terrestris and Cinnamomum zeylanicum extracts and trehalose added to diluents on goat epididymal sperm freezability.
This study investigates the effect of adding Tribulus terrestris ethanol extract (TEE) and Cinnamomum zeylanicum ethanol extract (CEE) and trehalose on freezability of goat epididymal spermatozoa. In Experiment 1, the treatments consist of basic extender containing 25, 50 or 100 μg/ml of TEE or CEE. The control contained no additives. Experiment 2 was carried out to compare the effect of best concentrations resulted in the first experiment with 150 mM trehalose added to basic extender. The results of experiment 1 showed that supplementation of 50 μg/ml TEE and 50 μg/ml CEE increased significantly the percentages o...
Source: Cryobiology - November 24, 2020 Category: Biology Authors: Ariyan F, Farshad A, Rostamzadeh J Tags: Cryobiology Source Type: research

Lipidomic changes in mouse oocytes vitrified in PEG 8000-supplemented vitrification solutions.
In this study, we examined whether supplementation of polyethylene glycol 8000 (PEG 8000) during vitrification influences the lipidome of the oocytes. We used liquid chromatography with tandem mass spectrometry (LC-MS/MS) to study the alteration in the lipidome in three groups of mouse oocytes: fresh, vitrified-warmed, and vitrified with PEG 8000-warmed during vitrification. In these groups, we targeted to analyze 21 lipid classes. We profiled 132 lipid species in the oocytes and statistical analyses revealed lipid classes that were up- or downregulated in these groups. Overall, our data revealed that several classes of li...
Source: Cryobiology - November 23, 2020 Category: Biology Authors: Jung GT, Lee J, Park D, Ahn JM, Um DE, Shin H, Lee JW, Kim J, Song H, Kim KP, Lim HJ Tags: Cryobiology Source Type: research

Addition of antifreeze protein type I or III to extenders for ram sperm cryopreservation.
In conclusion, the use of AFP, predominantly type I, may increase sperm cell protection during cryopreservation, with no adverse effect on potential fertilization capacity or increase in reactive oxygen species, being a potential cryoprotectant to ram sperm. PMID: 33186590 [PubMed - as supplied by publisher] (Source: Cryobiology)
Source: Cryobiology - November 10, 2020 Category: Biology Authors: Correia LFL, Espírito-Santo CG, Braga RF, Carvalho-de-Paula CJ, da Silva AA, Brandão FZ, Freitas VJF, Ungerfeld R, Souza-Fabjan JMG Tags: Cryobiology Source Type: research

Optimizing the spray parameters of a cryospray process.
Abstract Smaller spray zone of single-hole nozzle (SHN) constrains cryospray as a treatment method suitable for lesions having diameter larger than 15 mm on the skin surface. The present study is an attempt to resolve this issue, through the improvement in conventional technique of spraying liquid nitrogen on cancerous lesion. A multi-hole nozzle (MHN) with 5 holes is fabricated to demarcate the variation in outcome when cryogen is sprayed through customised MHN instead of conventional SHN. Special emphasis is placed on reducing the number of sitting required for completion of treatment and increasing the feasib...
Source: Cryobiology - November 7, 2020 Category: Biology Authors: Srivastava P, Kumar A Tags: Cryobiology Source Type: research

Ovary cold storage and shipment affect oocyte yield and cleavage rate of cat immature vitrified oocytes.
This study employed ovariectomy-derived domestic cat gonads as a model for wild felids, and aimed to compare the yield and developmental competence of Cryotop-vitrified oocytes (VOs) collected and cryopreserved right after ovary excision (In loco-VOs) or after 24 h cooled transport of ovaries (Shipped-VOs). The number of collected oocytes was higher in In loco-VOs than in Shipped-VOs (mean ± SD: 8 ± 3.36 vs 5.6 ± 3.1, p = 0.05). In vitro embryo production resulted in similar maturation (35% for both vitrified groups, p = 1) and fertilization rates (In loco-VOs: 29.1%; Shipped-VOs:...
Source: Cryobiology - November 7, 2020 Category: Biology Authors: Colombo M, Zahmel J, Binder C, Herbel J, Luvoni GC, Jewgenow K Tags: Cryobiology Source Type: research

Rapid quantification of multi-cryoprotectant toxicity using an automated liquid handling method.
Abstract Cryopreservation in a vitrified state has vast potential for long-term storage of tissues and organs that may be damaged by ice formation. However, the toxicity imparted by the high concentration of cryoprotectants (CPAs) required to vitrify these specimens remains a hurdle. To address this challenge, we previously developed a mathematical approach to design less toxic CPA equilibration methods based on the minimization of a toxicity cost function. This approach was used to design improved methods for equilibration of bovine pulmonary artery endothelial cells (BPAEC) with glycerol. To fully capitalize on ...
Source: Cryobiology - November 3, 2020 Category: Biology Authors: Warner RM, Ampo E, Nelson D, Benson JD, Eroglu A, Higgins AZ Tags: Cryobiology Source Type: research

Supplementation of ram's semen extender with Mito-TEMPO I: Improvement in quality parameters and reproductive performance of cooled-stored semen.
In this study, diluted semen samples were assigned into 5 parts, and received 0, 0.5, 5, 50 and 500 μM Mito-TEMPO. The prepared samples were stored at 5 °C up to 48 h. Chilled sperm motility, viability, abnormal morphology, mitochondrial membrane potential, membrane functionality and malondialdehyde concentration were assessed during 0, 24 and 48 h. For evaluation of reproductive performance, artificial insemination was performed via 24 h-chilled semen. In results, at time 0, no difference was observed among groups. Using 5 and 50 μM Mito-TEMPO resulted in higher (P ≤ 0.05) cooled sperm total mot...
Source: Cryobiology - November 2, 2020 Category: Biology Authors: Zarei F, Kia HD, Masoudi R, Moghaddam G, Ebrahimi M Tags: Cryobiology Source Type: research

Cryopreservation of human induced pluripotent stem cells by using a new CryoLogic vitrification method.
In this study, we developed a new friendly CVM by loading a small piece of hiPSCs colonies in the vitrification solution to the hook of Fiberplug to increase the cooling rate. To warm them, the CVM Fiberplug was immersed directly in a 37 °C warming solution for 1 min, and hiPSCs were then transferred to mTeSR1 medium. The result revealed that the new CVM had a high recovery rate and maintained the stemness and differentiation potential of hiPSCs. Our new CVM not only provide a safe way for hiPSCs preservation but also has a high survival rate in the storage of hiPSCs. PMID: 33147488 [PubMed - as supplied by pu...
Source: Cryobiology - November 1, 2020 Category: Biology Authors: Zhang Y, Liu H, Liu Z, Long P, Zhao X, Li Q, Huang Y, Ma Y Tags: Cryobiology Source Type: research

Low cryoprotectant concentration rapid vitrification of mouse oocytes and embryos.
In conclusion, the LCSM protocol, with one medium of much lower cryoprotectant concentrations and shorter equilibration time, reduces exposure to cryoprotectant toxicity while improves efficiency, consistency and reliability for mammalian oocyte and embryo preservation. PMID: 33137307 [PubMed - as supplied by publisher] (Source: Cryobiology)
Source: Cryobiology - October 30, 2020 Category: Biology Authors: Liu J, Lee GY, Biggers JD, Toth TL, Toner M Tags: Cryobiology Source Type: research

Oxidative and nitrosative stress in frozen-thawed pig spermatozoa. II: Effect of the addition of saccharides to freezing medium on sperm function.
ute;nchez R Abstract Saccharides have bioprotective properties, with a high capacity to preserve biological proteins and membranes during sperm cryopreservation. The aim of this study was to evaluate how replacing the lactose of cryopreservation media by sucrose (SUC) or trehalose (TRE) at concentrations of 0.2 M (SUC-1 and TRE-1) and 0.25 M (SUC-2 and TRE-2) affects frozen/thawed pig spermatozoa. The media used were composed of medium A (saccharide/egg yolk) and B (saccharide/egg yolk/glycerol), their osmolality being determined prior to freezing. Cell viability, membrane lipid disorder, acrosome integr...
Source: Cryobiology - October 27, 2020 Category: Biology Authors: Pezo F, Zambrano F, Uribe P, Risopatrón J, Moya C, Cesar de Andrade AF, Burgos RA, Yeste M, Sánchez R Tags: Cryobiology Source Type: research

Effect of Simmental bull seminal plasma protein in egg yolk-citrate extender on Kacang buck semen fertility.
The objectives of this study were to determine whether the supplementation of superior Simmental bull seminal plasma protein in egg yolk-citrate extender could improve the quality of post-thawed Kacang buck sperm, increase conceptions rates, and improve kidding rates. Buck semen was diluted without supplementation (T0) and with supplementation of 2.5 mg (T1) and 5 mg (T2) of Simmental bull seminal plasma protein per mL egg yolk-citrate extender. Extended semen was packed in 0.25 mL straw as cryopreserved frozen semen. Post-thawed semen samples were evaluated for viability, motility, intact plasma membranes, ...
Source: Cryobiology - October 22, 2020 Category: Biology Authors: Susilowati S, Triana IN, Sardjito T, Suprayogi TW, Wurlina W, Mustofa I Tags: Cryobiology Source Type: research

Regulation of NF- κB, FHC and SOD2 in response to oxidative stress in the freeze tolerant wood frog, Rana sylvatica.
Regulation of NF-κB, FHC and SOD2 in response to oxidative stress in the freeze tolerant wood frog, Rana sylvatica. Cryobiology. 2020 Oct 17;: Authors: Gupta A, Brooks C, Storey KB Abstract The wood frog, Rana sylvatica, is the primary model animal used for studying vertebrate freeze tolerance. Freeze tolerance adaptations by wood frogs are usually mediated by a set of well-tuned regulatory controls at the molecular level, starting from cell signal transduction and gene expression events that are ultimately reflected in protective responses by multiple cell systems. Previous studies provided exc...
Source: Cryobiology - October 17, 2020 Category: Biology Authors: Gupta A, Brooks C, Storey KB Tags: Cryobiology Source Type: research

The proteomic characterization of ram sperm during cryopreservation analyzed by the two-dimensional electrophoresis coupled with mass spectrometry.
Abstract The aim of this study was to analyze the effects of the cryopreservation process on the protein profile of ram sperm using two-dimensional electrophoresis (2-DE) coupled with mass spectroscopy. Semen was collected from five rams and cryopreserved in a Tris-based extender supplemented with glycerol and egg yolk as the main cryoprotectants. The fresh and post-thaw sperm total proteins were extracted and purified, followed by the 2-DE. The differential proteins in the stained gel were determined by mass spectrometry. The results indicated that there were 39 differential proteins between fresh sperm and froze...
Source: Cryobiology - October 14, 2020 Category: Biology Authors: Lv C, Larbi A, Memon S, Liang J, Zhao X, Shao Q, Wu G, Quan G Tags: Cryobiology Source Type: research

Successful vitrification of early-stage porcine cloned embryos.
In conclusion, our data demonstrate that the early-stage porcine cloned embryos including zygotes, 2- and 4-cells can be successfully vitrified, with respectable blastocyst yield and quality. PMID: 33065107 [PubMed - as supplied by publisher] (Source: Cryobiology)
Source: Cryobiology - October 13, 2020 Category: Biology Authors: Jia B, Xiang D, Guo J, Jiao D, Quan G, Hong Q, Fu X, Wei H, Wu G Tags: Cryobiology Source Type: research

Development of a spermatogonia cryopreservation protocol for blue catfish, Ictalurus furcatus.
Abstract Sustainability of channel catfish, Ictalurus punctatus ♀ × blue catfish, Ictalurus furcatus ♂ hybrid aquaculture relies on new innovative technologies to maximize fry output. Transplanting spermatogonia from blue catfish into channel catfish hosts has the potential to greatly increase gamete availability, and cryopreservation would make these cells readily accessible year-round. However, a protocol for cryopreserving blue catfish testicular tissues has not yet been fully developed. Therefore, the objectives of this experiment were to identify the best permeating [dimethyl sulfoxide (DMSO), e...
Source: Cryobiology - October 12, 2020 Category: Biology Authors: Abualreesh M, Myers JN, Gurbatow J, Johnson A, Xing, Wang J, Li S, Coogan M, Vo K, El Husseini N, Dunham RA, Butts IAE Tags: Cryobiology Source Type: research

Impact of embryo technologies on secondary sex ratio in rabbit.
In conclusion, we show that the MOVET program causes a direct, accumulative and long-term transmission of skewed SSR. PMID: 33053364 [PubMed - as supplied by publisher] (Source: Cryobiology)
Source: Cryobiology - October 11, 2020 Category: Biology Authors: Garcia-Dominguez X, Juarez JD, Vicente JS, Marco-Jiménez F Tags: Cryobiology Source Type: research

Addition of butylated hydroxytoluene (BHT) in tris-based extender improves post-thaw quality and motion dynamics of dog spermatozoa.
Abstract The aim of this study was to evaluate the interaction of different concentrations of butylated hydroxytoluene (BHT) in a tris-based extender on semen quality parameters in post-thawed dog semen. Twenty-four ejaculates were collected from eight male Beagle dogs using an artificial vagina. Pooled semen was diluted with a tris-based extender supplemented with 0 (control), 0.5, 1.0, 1.5, and 2.0 mM BHT, at a final concentration of 200 × 106 spermatozoa/mL. After thawing, sperm samples were assessed for motility parameters (CASA), membrane integrity (SYBR-14/PI), acrosome integrity (FITC-PNA), mito...
Source: Cryobiology - October 8, 2020 Category: Biology Authors: Sun L, Wu C, Xu J, Zhang S, Dai J, Zhang D Tags: Cryobiology Source Type: research

Exploring the role of E coli derived enzyme, oxyrase, as an oxygen scavanger to improve the cryotolerance of spermatozoa of sahiwal bull.
Abstract The current study intended to optimize the concentration of Oxyrase in the semen dilutor and to evaluate its effect on freezability of spermatozoa of Sahiwal bulls. Supplementation of Oxyrase at 0.125 IU/mL concentration significantly reduced dissolved oxygen (DO) in the dilutor to 4 ppm in 16-18 min at 35 °C. For supplementaion studies, a total of 24 eajaculates were categorized into poor and good ejaculates categories (n = 12 each) based on their initial progressive motility. Each ejaculate was further divided into two aliquotes. The first aliquote was diluted with tris-egg yolk extender w...
Source: Cryobiology - October 7, 2020 Category: Biology Authors: Ngou AA, Ghosh SK, Prasad JK, Katiyar R, Kumar A, Rautela R, Amarjeet, Srivastava N, Kumar A Tags: Cryobiology Source Type: research

Optimization of triacetate cellulose hollow fiber vitrification (HFV) method for cryopreservation of in vitro matured bovine oocytes.
Abstract The aim of the current work was to evaluate applicability of triacetate cellulose hollow fiber vitrification (HFV) method for cryopreservation of groups of in vitro matured bovine oocytes (12-17 oocytes per device). We also attempted to optimize HFV protocol by altering concentration of non-permeating cryoprotectant (sucrose) in vitrification solution and concentration of extracellular Ca2+ by using a calcium-free base medium for preparation of vitrification/rewarming solutions with ethylene glycol (EG) as a single permeating cryoprotectant. Neither of modifications of HFV protocol significantly affected ...
Source: Cryobiology - October 7, 2020 Category: Biology Authors: Kornienko EV, Romanova AB, Ikonopistseva MV, Malenko GP Tags: Cryobiology Source Type: research

Skim milk powder used as a non-permeable cryoprotectant reduces oxidative and DNA damage in cryopreserved zebrafish sperm.
Abstract Cryoprotectants play a vital role in the cryopreservation process, protecting biological samples from freezing damage. Here, we evaluate the effects of the combination and interaction of different extenders with permeable and non-permeable cryoprotectants, on the cryopreservation of Danio rerio sperm, analyzing the effects of cryopreservation through a broad approach to variables. Two extenders were used, Hank's balanced salt solution (HBSS) and Ginsburg's solution. Eight cryoprotective solutions (CS) were used: CS1 (HBSS + Me2SO 8%), CS2 (HBSS + Methanol 8%), CS3 (HBSS + Me2...
Source: Cryobiology - October 7, 2020 Category: Biology Authors: Rodrigues RB, Uczay M, Brito VB, Nunes Fossati AA, Godoy AC, Moura DJ, Ganz Vogel CI, Nogueira Vasconcelos AC, Streit DP Tags: Cryobiology Source Type: research

Carboxylated ε-poly-L-lysine, a cryoprotective agent, is an effective partner of ethylene glycol for the vitrification of embryos at various preimplantation stages.
In this study, we developed a common vitrification protocol using carboxlated ε-poly-l-lysine (COOH-PLL), a new cryoprotective agent for the vitrification of mouse embryos at different stages. The IVF-derived Crl:CD1(ICR) x B6D2F1/Crl pronuclear, 2-cell, 4-cell, and 8-cell, morula and blastocyst stage embryos were vitrified with 15% (v/v) ethylene glycol (EG) and 10% (w/v) COOH-PLL (E15P15) or 15% (v/v) EG and 15% (v/v) dimethyl sulfoxide (E15D15) using the minimal volume cooling method. The survival of vitrified embryos from pronuclear to blastocyst stages was equivalent between E15P15 and E15D15 groups. However, ...
Source: Cryobiology - October 6, 2020 Category: Biology Authors: Kawasaki Y, Kohaya N, Shibao Y, Suyama A, Kageyama A, Fujiwara K, Kamoshita M, Matsumura K, Hyon SH, Ito J, Kashiwazaki N Tags: Cryobiology Source Type: research

The efficacy of ice recrystallization inhibitors in rat lung cryopreservation using a low cost technique for ex vivo subnormothermic lung perfusion.
Abstract Although lung transplant remains the only option for patients with end-stage lung failure, short preservation times result in an inability to meet patient demand. Successful cryopreservation may ameliorate this problem; however, very little research has been performed on lung cryopreservation due to the inability to prevent ice nucleation or growth. Therefore, this research sought to characterize the efficacy of small-molecule ice recrystallization inhibitors (IRIs) for lung cryopreservation given their well-documented ability to control ice growth. Sprague-Dawley heart-lung blocks were perfused at room t...
Source: Cryobiology - October 5, 2020 Category: Biology Authors: Lautner L, Himmat S, Acker JP, Nagendran J Tags: Cryobiology Source Type: research