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Enrique Dupré, Javiera Carvajal
The natural population of the edible red sea urchin, Loxechinus albus, is decreasing due to overfishing. The embryos and larvae of the species are highly useful for monitoring marine pollution, which makes it necessary to conserve gametes, embryos and larvae to facilitate their use in diverse areas of aquaculture and environmental quality monitoring. This need can be met by cryopreserving individuals representing the different developmental stages to provide an ongoing supply of genetic material of the species...
November 23, 2018: Cryobiology
Liangpeng Ge, Songquan Yang, Hao Liang, Xueqin Liu, Wen Liu, Yuchun Ding, Yong Huang, Zuohua Liu
At refrigeration temperature, mouse embryos can retain their developmental ability for a couple of days. Previous research reports have focused on the effect of cool temperature on the development of 2-cell stage embryos, morulae or blastocysts and determined that the embryo still has the ability to produce offspring after about 48 h storage at refrigeration temperature. Here we examined whether refrigeration temperature affects the development of the eight-cell stage and if the stored eight-cell stage embryo can still be used as a host embryo for ES cell injection...
November 22, 2018: Cryobiology
James R Cypser, Wallace S Chick, Gregory M Fahy, Garrett J Schumacher, Thomas E Johnson
We report here a new, unbiased forward genetic method that uses transposon-mediated mutagenesis to enable the identification of mutations that confer cryoprotectant toxicity resistance (CTR). Our method is to select for resistance to the toxic effects of M22, a much-studied whole-organ vitrification solution. We report finding and characterizing six mutants that are resistant to M22. These mutants fall into six independent biochemical pathways not previously linked to cryoprotectant toxicity (CT). The genes associated with the mutations were Gm14005, Myh9, Nrg2, Pura, Fgd2, Pim1, Opa1, Hes1, Hsbp1, and Ywhag...
November 17, 2018: Cryobiology
Bin Wang, Gaoli Liu, Vasudevan Balamurugan, Yulong Sui, Guannan Wang, Yisheng Song, Qing Chang
Cryopreservation of tissue cells is an important method to maintain cell viability and cellular function. However, cell viability and function are less than ideal by conventional cell cryopreservation methods, which may result in apoptosis and necrosis of cells in cryopreservation. Trehalose plays a role in maintaining cell structure and protecting cells from stress. However, owing to the difficulty in transport of trehalose across the cell membrane, its antifreeze effect is limited. A large amount of trehalose (up to 237 ± 8...
November 17, 2018: Cryobiology
Deniz Tiryaki, İhsan Aydın, Ökkeş Atıcı
We have isolated psychrotolerant bacteria from the leaf apoplast of cold-adapted wild plants and aimed to investigate their effect on the cold resistance of bean (Phaseolus vulgaris L.). Based on the findings of 16S rRNA gene sequence analysis, 20 isolates belonging to 5 bacteria species (Pseudomonas fragi, P. chloropaphis, P. fluorescens, P. proteolytica and Brevibacterium frigoritolerans) were identified in the leaf apoplastic fluid of Draba nemorosa, Galanthus gracilis, Colchicum speciousum, Scilla siberica, Erodium cicutarium, respectively...
November 9, 2018: Cryobiology
Rodrigo Freitas Bittencourt, Eunice Oba, Carmo Emanuel de Almeida Biscarde, Hymerson Costa Azevedo, Marta Vasconcelos Bittencourt, Gabriel Felipe Oliveira de Menezes, Adrielle da Silva Lima, Kárita da Mata Fuchs, Antônio de Lisboa Ribeiro Filho
Eighteen semen samples from nine Santa Ines rams were cryopreserved in order to study the cryoprotective capacity of dimethylacetamide (DMA) at two concentrations (3% and 6%), and its interaction with trehalose (TRE, 100 mOsmol). A further objective of this study was to evaluate in vivo fertility of ram semen cryopreserved with dimethylacetamide. The control treatment was an extender medium with 6% glycerol (GLY). Five treatments were examined: 6% GLY, 3% DMA, 6% DMA, 3% DMA + TRE, and 6% DMA + TRE. After thawing, the kinetic sperm parameters were analyzed using a computerized system...
November 1, 2018: Cryobiology
Lei Yang, Lanjie Lei, Qihan Zhao, Zhen Gao, Xiaoyuan Xu
The cryopreservation of cumulus oocyte complexes (COCs) is an important process in reproductive medicine. However, this process slows both embryo and offspring development. In this study, the effects of maturation medium supplementation with lycium barbarum polysaccharide (LBP) on the developmental competence of vitrified-warmed mouse COCs and the possible mechanisms facilitating this process were investigated. The addition of LBP to the maturation medium improved the maturation rate and enhanced the developmental competence of vitrified-warmed mouse COCs...
October 31, 2018: Cryobiology
Mary Hagedorn, Zoltan Varga, Ronald B Walter, Terrence R Tiersch
The genetic resources of aquatic biomedical model organisms are the products of millions of years of evolution, decades of scientific development, and hundreds of millions of dollars of research funding investment. Genetic resources (e.g., specific alleles, transgenes, or combinations) of each model organism can be considered a form of scientific wealth that can be accumulated and exchanged, typically in the form of live animals or germplasm. Large-scale maintenance of live aquatic organisms that carry these genetic resources is inefficient, costly, and risky...
October 31, 2018: Cryobiology
Yi Yang, Wei-Wei Ma, Min-Wei Zhou, Zong-You Chen, Jian-Bin Xiang, Zhen-Yang Li, Bo Sun, Xiao-Dong Gu
OBJECTIVES: Peritoneal carcinomatosis is one of the causes of death in patients with advanced gastric cancer. We assumed that cryoablation could be applied as adjuvant therapy to control peritoneal carcinomatosis from gastric cancer. METHODS: We investigated the feasibility of cryoablation technique in rabbit model using a novel cryoablation balloon probe. The cryozones were harvested 7 days after cryoablation for histological evaluation. The levels of cytokines in the peripheral blood of rabbits were also detected...
October 27, 2018: Cryobiology
Matthew J Powell-Palm, Jessica Preciado, Chenang Lyu, Boris Rubinsky
In comparison with isobaric (constant pressure) freezing, isochoric (constant volume) freezing reduces potential mechanical damage from ice crystals and exposes stored biological matter to a lower extracellular concentration, at the price of increased hydrostatic pressure. This study evaluates the effects of isochoric freezing to low temperatures and high pressures on Escherichia coli (E. coli) survival. The viability of E. coli was examined after freezing to final temperatures between -5 °C and -20 °C for periods from 0...
October 23, 2018: Cryobiology
Md Ariful Alam, Sheikh Mustafizur Rahman, Yoji Yamamoto, Ricardo Shohei Hattori, Toru Suzuki, Manabu Watanabe, Carlos Augusto Strüssmann
Microinjection has proven useful for introduction of low-permeability cryoprotective agents (CPAs) into fish eggs or embryos for cryopreservation. In this work, we examined the suitable conditions for single or combined microinjection into the perivitelline space (PS) and the yolk mass (YM) of embryos of the Japanese whiting, an alternative marine fish model for embryo cryopreservation studies. The parameters examined were injection volume, CPA type and concentration, vehicle (diluent), and suitable developmental stage...
October 9, 2018: Cryobiology
Taylor Rutt, Nasim Eskandari, Maria Zhurova, Janet A W Elliott, Locksley E McGann, Jason P Acker, John A Nychka
Despite success in cryopreservation of cells in suspension, cryopreservation of cells in monolayers is still challenging. One of the major problems is detachment of the cells from the substrate which occurs during cryopreservation. We hypothesized that this detachment may be due to a mismatch in the coefficient of linear thermal expansion (αL ) between glass and the frozen cell layer which manifests as residual stress and stress relaxation. This mismatch results in a difference between the thermal expansion of ice and glass as they undergo temperature changes...
October 9, 2018: Cryobiology
Sanghwa Lee, Jun Hyuck Lee, Han-Woo Kim, Jong Wook Hong
Ice binding proteins (IBPs) have been attracting significant interest on account of their characteristic of inhibiting ice growth and recrystallization. Owing to their unique characteristics, IBPs have been studied for applications in food, pharmaceuticals, and medicine, as well as from a general scientific point of view. In this study, we have used differential scanning calorimetry (DSC) and Raman spectroscopy as tools to understand the ice binding activity of the Arctic-yeast-originating extracellular ice binding glycoprotein (LeIBP) isolated from Leucosporidium sp...
October 5, 2018: Cryobiology
Camila Knecht, Cecilia L Balaban, Joaquín V Rodríguez, Eduardo A Ceccarelli, Edgardo E Guibert, Germán L Rosano
Cold storage is a common procedure for liver preservation in a transplant setting. However, during cold ischemia, the liver suffers molecular alterations that can affect its performance. Also, deleterious mechanisms set forth in the storage phase are exacerbated during reperfusion. This study aimed to identify liver proteins associated with injury during cold storage and/or normothermic reperfusion using the isolated perfused rat liver model. Livers from male rats were subjected to either (1) cold storage for 24 h, (2) ex vivo normothermic reperfusion for 90 min or (3) cold storage for 24 h followed by ex vivo normothermic reperfusion for 90 min...
October 5, 2018: Cryobiology
J Branchereau, K Renaudin, Delphine Kervella, S Bernadet, Georges Karam, Gilles Blancho, D Cantarovich
BACKGROUND: There are currently two approaches to hypothermic preservation for most solid organs: static or dynamic. Cold storage is the main method used for static storage (SS), while hypothermic pulsatile perfusion (HPP) and other machine perfusion-based methods, such as normothermic machine perfusion and oxygen persufflation, are the methods used for dynamic preservation. HPP is currently approved for kidney transplantation. METHODS: We evaluated, for the first time, the feasibility of HPP on 11 human pancreases contraindicated for clinical transplantation because of advanced age and/or history of severe alcoholism and/or abnormal laboratory tests...
October 4, 2018: Cryobiology
Nasim Eskandari, Leah A Marquez-Curtis, Locksley E McGann, Janet A W Elliott
Cryopreservation of endothelium is one of the major challenges in the cryopreservation of complex tissues. Human umbilical vein endothelial cells (HUVECs) in suspension are available commercially and recently their post-thaw cell membrane integrity was significantly improved by cryopreservation in 5% dimethyl sulfoxide (Me2 SO) and 6% hydroxyethyl starch (HES). However, cryopreservation of cells in monolayers has been elusive. The exact mechanisms of damage during cell monolayer cryopreservation are still under investigation...
October 4, 2018: Cryobiology
Alla Orekhova, Miloš Barták, Josef Hájek
The soil microalgae of the genus Heterococcus are found in cold environments and have been reported for the terrestrial ecosystems of several Sub-Antarctic and Antarctic Islands. This study focused on resistance of Heterococcus sp. to sub-zero temperature. Heterococcus sp. was isolated from soil samples from James Ross Island, Antarctica. Culture of Heterococcus sp. grown in liquid medium were used to study ribitol effects at sub-zero temperatures on the species resistance to rapid freezing (RF, immersion of a sample into liquid nitrogen) and consequent cultivation on agar...
October 4, 2018: Cryobiology
Maria Kokotsaki, Mario Mairhofer, Christian Schneeberger, Julian Marschalek, Detlef Pietrowski
INTRODUCTION: Cryopreservation of ovarian tissue is an essential step in Ovarian Tissue Banking. In order to prevent the formation of ice crystals, typically the tissue is slowly frozen using a cryoprotectant. As an alternative the method of ultra-fast freezing by vitrification becomes more attention for freezing ovarian tissue because it has successfully been used for oocytes, embryos and sperm. However the impact of vitrification on granulosa cells, which are an essential part of ovarian tissue is uncertain...
September 25, 2018: Cryobiology
Claudia Köpnick, Marion Grübe, Johanna Stock, Angelika Senula, Hans-Peter Mock, Manuela Nagel
The potato's great genetic diversity needs to be maintained for future agricultural applications and can be preserved at ultra-low temperatures. To decipher detailed physiological processes, the aim of the study was to analyze the regrowth in 28 gene bank accessions and to reveal metabolite changes in a subset of four accessions that showed pronounced differences after shoot tip cryopreservation using DMSO droplet freezing and PVS3 droplet vitrification. Regrowth varied in all 28 genotypes ranging from 5% ('Kagiri') to 100% ('Karakter') and was higher after PVS3 droplet vitrification (71 ± 19%) than after cryopreservation using DMSO (54 ± 17%)...
September 23, 2018: Cryobiology
Momoh Karmah Mbogba, Zeeshan Haider, S M Chapal Hossain, Daobin Huang, Kashan Memon, Fazil Panhwar, Zeling Lei, Gang Zhao
For most of the cells, water permeability and plasma membrane properties play a vital role in the optimal protocol for successful cryopreservation. Measuring the water permeability of cells during subzero temperature is essential. So far, there is no perfect segmentation technique to be used for the image processing task on subzero temperature accurately. The ice formation and variable background during freezing posed a significant challenge for most of the conventional segmentation algorithms. Thus, a robust and accurate segmentation approach that can accurately extract cells from extracellular ice that surrounding the cell boundary is needed...
September 13, 2018: Cryobiology
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