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Lei Li, Zhongrong Chen, Mingke Zhang, Fazil Panhwar, Cai Gao, Gang Zhao, Bo Jin, Bin Ye
HepG2 cells have a number of research applications and cryopreservation of these cells would improve supply and thus facilitate the study. Development of effective cryopreservation protocols relies on knowledges of the fundamental mass transport characteristics of HepG2 cell membrane. Currently, the permeability parameters estimated from single-step addition are routinely used to predict the osmotic responses of the cells in multistep protocols, as well as used for prediction of optimal cooling rates. However, the reasonability of this approach has not been rigorously studied...
October 5, 2017: Cryobiology
Azam Dalman, Nafiseh Sadat Deheshkar Gooneh Farahani, Mehdi Totonchi, Reihaneh Pirjani, Bita Ebrahimi, Mojtaba Rezazadeh Valojerdi
This study compared slow freezing and vitrification of ovarian tissue by evaluation of histological changes, WNT signaling pathway and apoptotic genes expression. Ovarian tissue was obtained from women aging 27-38 years old. Ovarian cortex from each patient was divided into three pieces and randomly grouped as slow freezing, vitrification and control groups for investigation of WNT signaling gene expression and β-CATENIN presence as well as histological studies. The stromal structure of all ovaries were preserved...
October 4, 2017: Cryobiology
sAbdelRaouf M Ghallab, Abdallah M Shahat, Aya M Fadl, Mohamed M Ayoub, Adel R Moawad
The aim of the present study was to evaluate the effects of supplementation of semen extender with various non-enzymatic antioxidants on the quality of cooled or cryopreserved Arabian stallion spermatozoa. Semen collected from four pure Arabian stallions was centrifuged at 600g for 15 min. Spermatozoa were then diluted in INRA-82 extender supplemented with bovine serum albumin (BSA; 0, 10, 15 and 20 mg/mL) or trehalose (0, 75, 100 and 150 mM) or zinc sulphate (0, 100, 150 and 200 μM). The diluted semen was then either cooled at 5 °C or cryopreserved in 0...
October 4, 2017: Cryobiology
Mohamed Ali, Musa M Musa, Sulaiman Alfadul, K Al-Sobayel
A trial was conducted to check effect of adding gum Arabic (GA) instead of egg yolk (EY) as a cryoprotectant for stallion sperm. Two experiments were designed; experiment I tested adding 3 levels of nonheated GA (i.e., 3, 6 and 9 g/100 mL diluents) in HF-20 extender. However, in experiment II the same levels were tested except that GA was heated at 80 °C for 60 min. HF-20 containing 10% of EY was used as control. In experiment I, sperm frozen in HF-20 containing nonheated GA exhibited lower percentages of motile sperm, progressively motile sperm and sperm with intact plasma membranes, vitality rate, and acrosome integrity after cooling or after deep freezing...
October 3, 2017: Cryobiology
James D Benson, Adam Z Higgins, Kunjan Desai, Ali Eroglu
There is growing need for cryopreserved tissue samples that can be used in transplantation and regenerative medicine. While a number of specific tissue types have been successfully cryopreserved, this success is not general, and there is not a uniform approach to cryopreservation of arbitrary tissues. Additionally, while there are a number of long-established approaches towards optimizing cryoprotocols in single cell suspensions, and even plated cell monolayers, computational approaches in tissue cryopreservation have classically been limited to explanatory models...
September 28, 2017: Cryobiology
Yichi Zhang, Rasha Al-Attar, Kenneth B Storey
Rana sylvatica, known as the wood frog, can survive extremely cold temperatures during winter by undergoing full-body freezing, where it tolerates freezing of 65-70% of its total body water. During freezing, cellular dehydration decreases damage to the cell by preventing ice crystallization. Challenged with many stresses, these animals are forced to develop physiological adaptations to osmoregulation and osmoprotection that are necessary to ensure their survival. The purpose of this study was to elucidate a potential mechanism by which the transcription factor, NFAT5, regulates the expression of three osmoregulatory proteins (aldose reductase, SMIT, and BGT-1)...
September 22, 2017: Cryobiology
Behnaz Hatef, Amirhossein Taromchi, Reza Nejatbakhsh, Ahmad Farrokhi, Saeed Shokri
The destructive effects of sperm cryopreservation result in reduced sperm motility and increased apoptosis. Oocytes, endometrium, and follicular fluid express stromal cell-derived factor-1 alpha (SDF-1α) or C-X-C motif chemokine ligand 12 (CXCL12) while its specific receptor chemokine, CXC motif receptor 4 (CXCR4) is expressed in the head of sperm. SDF-1α can increase sperm motility and preserve normal mitochondrial status. The present study intends to investigate whether the addition of SDF-1α to freezing extender can facilitate cryosurvival of spermatozoa and how SDF-1α protects spermatozoa against damages during cryopreservation...
September 22, 2017: Cryobiology
Yuxin Zheng, Nina Zhang, Shujie Liu, Qingwang Li, Zhongliang Jiang
In this study, water-soluble Laminaria japonica polysaccharide3 (LJP-P3) was investigated for the cryoprotective effects on bull sperm. Five concentrations of LJP-P3 with 0.1, 1, 10, 50 and 100 mmol/L were added into the extenders of bull semen, respectively, and the effects on quality of sperm after freezing-thawing were assessed. The results showed that the kinematic parameters of bull sperm including linear motile sperm (LM), curvilinear line velocity (VCL) value, straight line velocity (VSL) and velocity of the average path (VAP) were greater in the extenders containing LJP-P3 (P<0...
September 20, 2017: Cryobiology
Valter Giaretto, Claudio Passerone
The paper presents a set of experiments that were performed to characterize the freezing front propagation in water first, and in an agar-gel solution afterwards. The experimental setup made of Peltier devices, to emulate the cryogenic effect, and a copper cold finger, to mimic the cold probe interface, are described. We claim that by monitoring some temperatures at the generating cryodevice, several pieces of information can be derived through the cold interface to assess the outside thermodynamic changes...
September 19, 2017: Cryobiology
Ying Luo, Peng Wang, Hui Liu, Zhengyan Zhu, Chenglong Li, Yingtang Gao
AIM: We aim to assess the effect of the state of T cells before cryopreservation on the post-thaw proliferative capacity, phenotype and functional response. METHODS: Peripheral blood mononuclear cells (PBMCs) were isolated from a hepatocellular carcinoma (HCC) patient, and the T cells were frozen during cell culture according to our experimental design. After a period of re-culture, the proliferative capacity of the cryopreserved cells, the expression of T cell surface markers and the secretion of IFN-γ and IL-10 were assayed...
August 31, 2017: Cryobiology
Hirokazu Kusakabe, Hiroyuki Tateno
In order to protect sperm chromosomes against degradation when they are being stored for relatively high temperatures, we investigated the optimal pH of the freeze-drying solution, EGTA/Tris-HCl buffered solution alkalinized by NaOH (Na-ETBS) or KOH (K-ETBS). Mouse spermatozoa suspended in Na-ETBS or K-ETBS were freeze-dried at pH 5.0-8.4 and stored at 4 °C or 50 °C for 3 days. Some freeze-dried samples were stored at 25 °C for 3 days or 1 month. After storage, samples injected into oocytes using intracytoplasmic sperm injection were assessed for chromosome damage in resulting zygotes...
August 30, 2017: Cryobiology
Farrokh Habibzadeh, Parham Habibzadeh, Jamshid Roozbeh
No abstract text is available yet for this article.
August 26, 2017: Cryobiology
Michelle H Connolly, Estefania Paredes, Peter Mazur
To date, traditional cryopreservation techniques have not been amenable to zebrafish (Danio rerio) embryos, due in part to their large yolky eggs, which have a low surface to volume ratio and limited permeability to water and cryoprotectants. However, recent vitrification and ultra-rapid warming studies in mice have demonstrated successful preservation by dehydrating 85% or more of their total water content. We hypothesized that this approach may help overcome the barriers to embryo cryopreservation among D...
August 12, 2017: Cryobiology
P C Dos Santos-Neto, F Cuadro, N Barrera, M Crispo, A Menchaca
The objective was to evaluate pregnancy outcomes and birth rate of in vivo derived vs. in vitro produced ovine embryos submitted to different cryopreservation methods. A total of 197 in vivo and 240 in vitro produced embryos were cryopreserved either by conventional freezing, or by vitrification with Cryotop or Spatula MVD methods on Day 6 after insemination/fertilization. After thawing/warming and transfer, embryo survival rate on Day 30 of gestation was affected by the source of the embryos (in vivo 53...
August 10, 2017: Cryobiology
Md Mahbubul Hassan, Xiaoxu Li, Jian G Qin
Low survival of cryopreserved sperm impedes the application of cryopreservation technique in spermcasting oyster species. This study developed a simple method of liquid nitrogen vapor freezing to improve post-thaw sperm survival in the spermcasting oyster Ostrea angasi. The results indicate that the permeable cryoprotectants, dimethyl sulfoxide (DMSO), ethylene glycol (EG) and propylene glycol (PG) were non-toxic to sperm up to 20% concentration and 90 min exposure whereas methanol at 10% or higher was toxic to sperm for any exposure over 30 min...
August 10, 2017: Cryobiology
Liana Monteiro da Fonseca Cardoso, Marcelo Alves Pinto, Andrea Henriques Pons, Luiz Anastácio Alves
Cryopreservation of hepatocytes is a crucial step in the implementation of cell therapy for treating certain liver diseases. In the present study we investigated the effect of the some disaccharides on the cryopreservation of rat hepatocytes. Liver cells were frozen in media in the presence or absence of low concentrations of Me2SO (5% Me2SO) supplemented with varying concentrations of disaccharides (sucrose, glucose and trehalose). After 7 days of cryopreservation, the hepatocytes were thawed and viability was measure by exclusion of trypan blue and by the MTT technique, as well as by determining albumin production...
August 4, 2017: Cryobiology
Franco Lugnani, Enric Gunther, Pedro Torrecillas, Carlos Galacho, Adolfo Jiménez Garrido, Paul Mikus, Nina Klein, Michael K Stehling, Matteo Macchioro, Liel Rubinsky, Narayan Raju, Boris Rubinsky
We report results from an acute, single case study in the pig liver on the effects of a tissue ablation protocol (we named cryoelectrolysis) in which 10 min of cryosurgery, with a commercial cryosurgical probe, are delivered after 10 min of electrolysis generated by a current of about 60 mA. The histological appearance of tissue treated with cryoelectrolysis is compared with the appearance of tissue treated with 10 min of cryosurgery alone and with 10 min of electrolysis alone. Histology done after 3 h survival shows that the mixed rim of live and dead cells found around the ablated lesion in both cryosurgery and electrolytic ablation is replaced by a sharp margin between life and dead cells in cryoelectrolysis...
August 3, 2017: Cryobiology
Kuerban Tusong, Xiaoxing Guo, Shanshan Meng, Xiaoning Liu, Ji Ma
The cold tolerance mechanisms of insect have been studied extensively on the model species Drosophila and a few other species at the transcriptional level. However studies on insects that inherit strong cold tolerance are limited. Cold hardy Tenebrionid beetle Microdera punctipennis is endemic to Gurbantonggut Desert, northwest of China. However, its genomic information is lacking. To investigate the overwintering mechanisms of M. punctipennis adult, RNA-seq was performed on the winter adults and the control adults that were kept in laboratory at 30 °C...
August 1, 2017: Cryobiology
Bushra Allah Rakha, Muhammad Sajjad Ansari, Shamim Akhter, Zartasha Zafar, Iftikhar Hussain, Julian Santiago-Moreno, Elisabeth Blesbois
The Indian red jungle fowl is a sub-species of the genus Gallus native to South Asia; facing high risk of extinction in its native habitat. During cryopreservation, permeable cryoprotectants like glycerol are usually employed and we previously showed encouraging results with 20% glycerol. Because bird spermatozoa contain very little intracellular water, the possibility of replacing an internal cryoprotectant by an external one is opened. In the present study, we tested the replacement of internal cryoprotectant glycerol by the external cryoprotectant Polyvinylpyrrolidone (PVP)...
July 27, 2017: Cryobiology
Mathieu Pasquier, Tomasz Darocha, Paul Husby
No abstract text is available yet for this article.
July 25, 2017: Cryobiology
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