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M Mahfujur Rahman, M Rahmat Ali, M Rafiqul Islam Sarder, M Fazlul Awal Mollah, Najmus Sakib Khan
The study was conducted to develop sperm cryopreservation protocol for the endangered spiny eel, Mastacembelus armatus as well as to use the cryopreserved sperm in breeding. Activation of sperm was evaluated by observing motility in different osmolalities of NaCl solution (48-319 mOsmol/kg) and found that sperm motility decreased with increasing osmolality of the NaCl solution and it was totally inhibited at 319 mOsmol/kg. The highest motility and swimming duration of sperm were observed as 14.5 ± 0.5 min and 4...
October 13, 2016: Cryobiology
Dowglish F Chaves, Iana S Campelo, Mirelly M A S Silva, Maajid H Bhat, Darcio I A Teixeira, Luciana M Melo, Joanna M G Souza-Fabjan, Pascal Mermillod, Vicente J F Freitas
The aim of this study was to evaluate the use of antifreeze protein type III (AFP III) into vitrification medium on meiotic spindle morphology of in vitro matured bovine oocytes as well as the fertilization and blastocyst rates. Mature cumulus-oocyte complexes (COC) were distributed in four groups: control (untreated), vitrified without supplementation (AFP0) or supplemented with 500 (AFP500) or 1000 ng/mL (AFP1000) into vitrification solutions. Samples from each group were used to analyze the organization of meiotic spindle by confocal microscopy and the remaining COC were submitted to in vitro fertilization and culture for eight days...
October 8, 2016: Cryobiology
Josef Hájek, Miloš Barták, Jana Hazdrová, Marie Forbelská
Extremophilic lichens and their photosynthesizing photobionts from the cold regions of Earth are adapted to perform photosynthesis at subzero temperatures. To evaluate interspecific differences in the critical temperature for primary photochemical processes of photosynthesis, we exposed lichen thalli of Usnea antarctica, Usnea aurantiaco-atra, and Umbilicaria cylindrica to linear cooling from +20 to -50 °C at a constant rate of 2 °C min(-1). Simultaneously, two chlorophyll fluorescence parameters (FV/FM - potential yield of photosynthetic processes in photosystem II, ΦPSII - effective quantum yield of PS II) evaluating a gradual subzero temperature-induced decline in photosynthetic processes were measured by a modulated fluorometer...
October 8, 2016: Cryobiology
Yue Zhang, Wei Li, Yongshun Ma, Dian Wang, Xiaoxue Zhao, Changjun Zeng, Ming Zhang, Xianyin Zeng, Qinggang Meng, Guangbin Zhou
The study was aimed to investigate the effect of melatonin on the development potential of mouse MII oocytes after cryopreservation. Mouse MII oocytes were subjected first to vitrification/warming and 2 h of in vitro culture (phase 1), then to parthenogenetic activation (PA) followed by in vitro culture of parthenogenetic embryos (phase 2). Different concentrations of melatonin (0, 10(-9), 10(-6) mol/L) were added to the medium during either phase 1, phase 2 or both phases. The fresh oocytes were used as control...
October 7, 2016: Cryobiology
Fabio Mosca, Manuela Madeddu, Ahmad Abdel Sayed, Luisa Zaniboni, Nicolaia Iaffaldano, Silvia Cerolini
The aim of this study was to assess the combined effect of dimethylacetamide (DMA) and two non-permeating cryoprotective agents, trehalose and sucrose, on the quality of post-thaw chicken semen. Adult Lohmann roosters (n = 27) were used. Semen was processed according to the following treatments: Lake pre-freezing extender +6% DMA (LPF, control treatment), LPF + 0.1 M trehalose (LPF-T treatment), LPF + 0.1 M sucrose (LPF-S treatment) and LPF + 0.1 M trehalose + 0.1 M sucrose (LPF-TS treatment)...
October 4, 2016: Cryobiology
Lavanya Pothana, Lalitha Devi, Naresh Kumar Venna, Niharika Pentakota, Vivek Phani Varma, Jedy Jose, Sandeep Goel
Cryopreservation of immature testis is a feasible approach for germplasm preservation of male animals. Combinations of dimethyl sulfoxide (DMSO) and foetal bovine serum (FBS) are used for testis cryopreservation. However, an alternative to FBS is needed, because FBS is expensive. Buffalo ocular fluid (BuOF), a slaughter house by-product, could be an economical option. The objective of the present study was to assess whether BuOF can replace FBS for cryopreservation of immature mouse (Mus musculus), rat (Rattus norvegicus), and buffalo (Bubalus bubalis) testes...
September 28, 2016: Cryobiology
A T M Cunha, J O Carvalho, N R Kussano, C F Martins, G B Mourão, M A N Dode
In this study we examined quality, longevity and ability of epididymal sperm (EP) to bind to oviduct explants (OE) after cooling and cryopreservation. Ejaculated (EJ) and EP sperm from seven bulls were evaluated before, during and after cryopreservation for total (TM), progressive motility (PM), sperm morphology, plasma membrane integrity (PMI) and acrosome integrity (ACI). For longevity, cryopreserved EP, EJ and a third group of cells in which EP spermatozoa were incubated with seminal plasm (SP) for 10 min after thawing (EPP group), were compared, and the groups were analyzed at 0, 3, 6, and 24 h for all parameters...
September 28, 2016: Cryobiology
Timothy J Morris, Andrew Picken, Duncan M C Sharp, Nigel K H Slater, Christopher J Hewitt, Karen Coopman
With the cell therapy industry continuing to grow, the ability to preserve clinical grade cells, including mesenchymal stem cells (MSCs), whilst retaining cell viability and function remains critical for the generation of off-the-shelf therapies. Cryopreservation of MSCs, using slow freezing, is an established process at lab scale. However, the cytotoxicity of cryoprotectants, like Me2SO, raises questions about the impact of prolonged cell exposure to cryoprotectant at temperatures >0 °C during processing of large cell batches for allogenic therapies prior to rapid cooling in a controlled rate freezer or in the clinic prior to administration...
September 20, 2016: Cryobiology
Takahiro Yamanaka, Kazuya Tashima, Rio Takahashi, Seiji Takashima, Teppei Goto, Masumi Hirabayashi, Shinichi Hochi
Two protocols, Bicell(®) freeze-thawing and Cryotop(®) vitrification-warming, were compared for suitability in cryopreserving rat pancreatic islets (101-150 μm in mean diameter). Immediate survival rates of post-thaw and post-warm islets (50 and 57%, respectively), assessed by FDA/PI double staining, were lower than that of fresh control islets (90%). Most of the PI-positive dead cells were detected in peripheral area of post-warm islets, and were removed after subsequent 24 h culture (survival rate; 85% vs 59% in post-thaw islets)...
September 17, 2016: Cryobiology
Jessye Wojtusik, Parker Pennington, Nucharin Songsasen, Luis R Padilla, Scott B Citino, Budhan S Pukazhenthi
Preserving genetic diversity of the critically endangered Addra gazelle (Nanger dama ruficollis) could be enhanced through the use of frozen-thawed sperm and artificial insemination. Our aim was to characterize Addra ejaculate traits and to assess the effects of cholesterol-loaded cyclodextrin (CLC) on sperm cryosurvival. Fresh ejaculates were treated with CLC (0, 0.5, 1.5, 3.0, and 6.0 mg/ml) prior to cryopreservation. All males produced spermic ejaculates with >75% sperm motility. The mean ± SEM seminal volume, sperm concentration, percent motility, forward progression, and percent morphologically normal spermatozoa were 3...
September 9, 2016: Cryobiology
Juliana P Alves, Carine D Corcini, Estela F Silva, Josie S Caldas, Tainã F Cardoso, Sergio R N Piedras, Rodrigo D Jardim, Antonio S Varela Junior
Amides were tested as internal cryoprotectants for the preservation of wild silverside (Odontesthes bonariensis) sperm. The semen was diluted in modified Mounib's medium and cryopreserved by adding 2, 5, 8 or 11% of dimethyl acetamide (DMA), dimethyl formamide (DMF) or methyl formamide (MF). Dimethyl sulfoxide (DMSO) at a concentration of 10% diluted in modified Mounib's medium was used as a control. The rate motility (17.7 ± 1.9%) and time motility (143.2 ± 9.7 s) (P < 0.05) of the sperm were higher with 2% DMF when compared with the other treatments...
September 5, 2016: Cryobiology
Samantha M Logan, Kenneth B Storey
Mammalian hibernation is characterized by a general suppression of energy expensive processes and a switch to lipid oxidation as the primary fuel source. Glucose-responsive carbohydrate responsive element binding protein (ChREBP) has yet to be studied in hibernating organisms, which prepare for the cold winter months by feeding until they exhibit an obesity-like state that is accompanied by naturally-induced and completely reversible insulin resistance. Studying ChREBP expression and activity in the hibernating 13-lined ground squirrel is important to better understand the molecular mechanisms that regulate energy metabolism under cellular stress...
October 2016: Cryobiology
Yichi Zhang, Shannon N Tessier, Kenneth B Storey
Foxo4 and MyoG proteins regulate the transcription of numerous genes, including the E3 ubiquitin ligases MAFbx and MuRF1, which are activated in skeletal muscle under atrophy-inducing conditions. In the thirteen-lined ground squirrel, there is little muscle wasting that occurs during hibernation, a process characterized by bouts of torpor and arousal, despite virtual inactivity. Consequently, we were interested in studying the regulatory role of Foxo4 and MyoG on ubiquitin ligases throughout torpor-arousal cycles...
October 2016: Cryobiology
Guangbin Zhou, Yan Zeng, Jiang Guo, Qinggang Meng, Qingyong Meng, Gongxue Jia, Keren Cheng, Changjun Zeng, Ming Zhang, Guoshi Liu, Shi'en Zhu
This study was conducted to determine the impact of vitrification on the expression of genes regulating pluripotency and apoptosis in mouse morulae. The morulae were randomly allocated into three groups: (1) untreated (control), (2) exposed to vitrification solution without freezing (toxicity), or (3) vitrified by open-pulled straw method (vitrification). In vitro development was evaluated by morphology and assessed by the blastocyst rate and the blastocyst total cell number. Gene expression in morulae and blastocysts was assessed by quantitative Real Time-PCR (qRT-PCR) and western blot...
October 2016: Cryobiology
Keiko Omori, Eiji Kobayashi, Jeffrey Rawson, Masafumi Takahashi, Yoko Mullen
Prolonged pancreas cold ischemia is known to negatively correlate with islet isolation outcomes, hindering successful islet transplantation to treat Type-1 Diabetes. Due to poor islet isolation outcome, pancreata with over 16 h cold ischemia are currently not considered for islet transplantation. Mechanisms involved in pancreas cold ischemia/rewarming mediated islet damage during islet isolation and culture are not well understood. Using an en bloc cold preserved rat pancreas preparation, we attempted to clarify possible mechanisms of islet death associated with prolonged pancreas cold ischemia and subsequent rewarming...
October 2016: Cryobiology
Y Horokhovatskyi, S Sampels, J Cosson, O Linhart, M Rodina, P Fedorov, M Blecha, B Dzyuba
The present study examined the lipid composition of plasma membranes in carp sperm with different post-thaw motility. The approach adapted for carp sperm cryopreservation, which involves the selection of the most effective protocol for individual males by comparing two cryoprotective media, was applied to the cryopreservation procedure. Sperm motility prior to freezing was greater than 80% but decreased to 40% in one group and to 10% in another group following cryopreservation. Lipid content of fresh sperm in all groups was analysed by thin layer chromatography and gas chromatography, with significant differences in phospholipid content, cholesterol and free fatty acids detected between groups, whereas the cholesterol/phospholipid ratio was extremely similar between groups (0...
October 2016: Cryobiology
Arash Valadbeygi, Tahere Naji, Afshin Pirnia, Mohammadreza Gholami
Successful freezed-thaw of adipose-derived mesenchymal stem cells (ADMSCs) could be a major step in regenerative medicine as well as in the cloning of animal breeds. The aim of this study was to evaluate the efficacy of selenium on the optimizing of freezed-thaw media in the ADMSCs. ADMSCs were extracted from NMRI mice and purified with positive selection Monoclonal CD105 Antibody (PE) and negative selection Monoclonal CD31 and CD45 Antibody using MACS method as well as differentiation to adipose and bone tissue...
October 2016: Cryobiology
Traimat Boonthai, Weerasith Khaopong, Jumlong Sangsong, Verapong Vuthiphandchai, Subuntith Nimrat
This objective of the study was to evaluate the effects of A. hydrophila subsp. hydrophila and P. fluorescens on sperm motility, sperm viability and sperm morphometry of cryopreserved silver barb (Barbodes gonionotus) semen and survival of tested bacteria after cryostorage. Semen was diluted in a calcium-free Hank's balanced salt solution (Ca-F HBSS) supplemented with or without 0.25% penicillin-streptomycin (PS) after which A. hydrophila subsp. hydrophila or P. fluorescens was immediately added into extended semen prior to freezing...
October 2016: Cryobiology
Timothy C Hawes
It has always been assumed that a three-dimensional protein structure is essential to antifreeze protein (AFP) ice interactions. Using a 9 kDa AFP isolated from the springtail, Gomphiocephalus hodgsoni, it was found that the bond between ice and protein is maintained independent of higher order protein structure. GomplyAFP9 remained bound to ice after denaturing by a range of agents (boiling, extreme pH, DTT, ethanol, urea). Thermal hysteresis was minimal (0.03-0.04 °C), but not lost. Crystal faceting and growth occurred normal to the c-axis, indicating the protein binds primarily to sites along the a-axis...
October 2016: Cryobiology
G A Bozhok, O S Sidorenko, E M Plaksina, T M Gurina, A N Sukach, V S Kholodnyy, V D Ustichenko, S B Bilyavskaya, T P Bondarenko, E I Legach
Stem/progenitor cells are thought to have the potential in the treatment of severe neurodegenerative diseases. Recently, sympathoadrenal progenitors expressing specific markers of neural crest derivatives and capable to differentiate into neurons were discovered in adult bovine and human adrenal glands, but there was no reported data on cryopreservation of sympathoadrenal progenitors. The aim of the present study was to examine the neural differentiation potential of sympathoadrenal progenitors derived from fresh and cryopreserved neonatal porcine adrenal glands...
October 2016: Cryobiology
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