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ABSTRACT ARCHIVE |
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CryoLetters is a bimonthly, international journal for low temperature science and technology |
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CryoLetters 41 (2), 57-61 (2020) Ice growth habits in solutions containing insect thermal hysteresis proteins compared to those with fish antifreeze proteins P. W. Wilson School of Environment, Science and Engineering, Southern Cross University, Lismore, NSW 2480, Australia Abstract BACKGROUND: The details of the mechanism of action of ice binding proteins (IBPs) have been intensively studied and hotly debated for some decades. OBJECTIVE: To outline the inherent differences between the manifested growth of single ice crystals grown in the presence of fish antifreeze proteins and those grown with insect thermal hysteresis proteins. MATERIALS AND METHODS: Observations of single ice crystals taking the shape of hexagonal bipyramids using a nanolitre osmometer and grown in the presence of so called antifreeze glycopeptides from the Antarctic fish species Dissostichus maswoni, are compared with those seen with insect thermal hysteresis proteins from Tenebrio molitor, grown in a Ramsay chamber, which grow as lemon-shaped crystals. RESULTS: The difference in growth allows us to infer methods of action of each class of protein. Further, below the thermal hysteresis gap, or non-equilibrium freezing point, the explosive growth seen with fish antifreeze proteins is demonstrated but is yet to be fully explored. CONCLUSION: Ice growth behaviour can be used to indicate or infer the crystal faces to which the molecules may have adsorbed. Keywords: antifreeze, fish, insect, hysteresis, ice binding.
CryoLetters 41 (2), 62-67 (2020) TREATMENT WITH MELATONIN ENHANCES THE EMBRYO QUALITY AND THE DEVELOPMENT OF VITRIFIED/WARMED EIGHT CELL MOUSE EMBRYOS BY SOLID SURFACE VITRIFICATION (SSV) Kubra Caglar1, Ahmet Kocabay2, Ali Cihan Taskin2* and Sezen Arat3 1 Department of Agricultural Biotechnology, Institute of Natural and Applied Sciences, Tekirdag Namik Kemal University, Tekirdag, Turkey. Abstract BACKGROUND: Melatonin is an endocrine hormone secreted from the pineal gland located outside the blood-brain barrier. OBJECTIVE: In this study, in vitro propagated eight-cell
mouse embryos were vitrified by the Solid Surface Vitrification (SSV) method and after thawing, their in vitro development and embryo qualities in melatonin added media were investigated. METHODS:
Pronuclear stage embryos obtained from super ovulated B6CBAF1/J strain mice, were cultured until the eight-cell stage. Then these eight-cell embryos
were vitrified by the SSV method and after thawing, cultured in melatonin added media at 37°C and 5 %CO2 conditions until the blastocyst stage. RESULT: In the experimental period, in vitro embryo development rates of the control, SSV and
Keywords: melatonin, vitrification, SSV, mouse, eight-cell stage, embryo development.
CryoLetters 41 (2), 68-74 (2020) EFFECT OF BUTYLATED HYDROXY TOLUENE AND VITAMIN E ON THE CRYOSURVIVABILITY OF BUCK SEMEN Raju Kr Dewry1*, BC Deka2, RK Biswas2, D Bhuyan2, Pranjal Borah2, 1 ICAR-National Dairy Research Institute, Karnal, Haryana-132001, India. Abstract BACKGROUND: The quality of frozen semen can be improved by supplementing Tris extender with antioxidant to prevent oxidation and maintain sperm motility. OBJECTIVE: To study the effects of adding combinations of suitable concentrations of butylated hydroxy toluene (BHT) and Vitamin E in Tris extender on the quality of frozen goat semen. MATERIALS AND METHODS : A total of 40 ejaculates collected from five Beetal bucks were used to study the effect on the quality of frozen semen of supplementing Tris extender with 200 µM BHT, 2 mM Vitamin E and 200 µM BHT + 2 mM Vitamin E. RESULTS: The sperm motility, live sperm, live intact acrosome and HOST-reacted sperm differed significantly (P<0.01) between stages and between antioxidants. There was no significant difference (P<0.05) in interaction between stages (equilibration, freezing) and antioxidants, except for HOST-reacted sperm. Critical difference test revealed that Tris extender containing 2 mM vitamin E showed significantly (P<0.05) higher sperm motility, live sperm, live intact acrosome and HOST-reacted sperm, and significantly (P<0.05) lower release of alanine transaminase (ALT) and aspartate transaminase (AST). CONCLUSION: Supplementation of Tris extender with 2 mM vitamin E maintained superior quality of frozen Beetal buck semen. Keywords: BHT, Frozen Buck semen, seminal attributes, vitamin E.
CryoLetters 41 (2), 75-85 (2020) CRYOPRESERVATION OF BLEEDING HEART (Lamprocapnos spectabilis (L.) FUKUHARA) SHOOT TIPS USING ENCAPSULATION-DEHYDRATION Dariusz Kulus UTP University of Science and Technology in Bydgoszcz, Faculty of Agriculture and Biotechnology, Laboratory of Ornamental Plants and Vegetable Crops, 6 Bernardyńska Str. 85-029 Bydgoszcz, Poland. Abstract BACKGROUND: Bleeding heart (Lamprocapnos spectabilis (L.) Fukuhara) is a popular vegetatively-propagated ornamental and medicinal plant species with recalcitrant seeds. Efforts have been made to increase the species diversity. However, there has been no research related to cryopreservation of any Fumariaceae species. OBJECTIVE: The aim of this study was to develop a cryopreservation protocol of Lamprocapnos spectabilis ‘White Gold’, and to evaluate genetic stability of the recovered plantlets. MATERIALS AND METHODS: Shoot tips precultured on Murashige and Skoog medium enriched with 9% sucrose and 10 µM abscisic acid were embedded in 3% calcium alginate, osmotically dehydrated in a sucrose gradient, and then air-desiccated for 0 to 5 h. After storing in liquid nitrogen (LN), the rewarmed shoot tips were placed on recovery media with various plant growth regulators. Morphometric analysis of the recovered shoots was performed. The genetic stability was evaluated using RAPD, ISSR and SCoT markers. RESULTS: The highest recovery of cryopreservation-derived explants (31.3 – 36.4%) was reported after 4-h desiccation and regrowth on cytokinin-supplemented medium. Shoots recovered from explants subjected to this cryoprocedure were longer and of higher fresh weight compared with the control. Cryopreservation also positively affected the further development of leaves. No genetic variation was detected by any of the three marker systems tested. CONCLUSION: Lamprocapnos spectabilis shoot tips have a limited dehydration tolerance. However, stress related to the cryoprocedure positively affects the development of shoots. Even though cryopreservation affects the viability of explants, it does not alter their DNA sequence. Keywords: desiccation, ISSR, morphometric analysis, RAPD, recovery medium, SCoT.
CryoLetters 41 (2), 86-91 (2020) IN VITRO DEVELOPMENT OF ZONA PELLUCIDA-FREE PORCINE ZYGOTES CULTURED INDIVIDUALLY AFTER VITRIFICATION Nhien Thi Nguyen1, Maki Hirata1*, Fuminori Tanihara1, Yoko Sato2, 1 Faculty of Bioscience and Bioindustry, Tokushima University, Tokushima 779-3233, Japan. Abstract BACKGROUND: Cryopreservation of zona pellucida (ZP)-free embryos provides more options for somatic cell nuclear transfer, particularly during handmade cloning. OBJECTIVE: This study investigated whether the removal of the ZP affects the development of porcine zygotes after vitrification and warming. MATERIALS AND METHODS: We determined the appropriate volume of the corresponding medium for the individual culture of ZP-intact and -free embryos and evaluated the protection effect of ZP during cryopreservation on the resulting development of the vitrified-warmed zygotes. RESULTS: The volume of culture medium influenced the development of ZP-intact zygotes, and a volume of 15 µL was most suitable for their development. However, the volume of culture medium did not modify the development of ZP-free zygotes. The removal of the ZP before vitrification did not adversely affect embryonic development or quality of the resulting blastocysts. CONCLUSION: Our results suggest that the removal of the ZP does not cause detrimental effects to the development of vitrified-warmed zygotes. Keywords: Cryotop, individual culture, porcine zygotes, vitrification, zona pellucida-free.
CryoLetters 41 (2), 92-99 (2020) EFFECT OF GLUTATHIONE SUPPLEMENTATION TO SEMEN EXTENDER ON POST-THAWED ROOSTER SPERM QUALITY INDICES FROZEN AFTER DIFFERENT EQUILIBRATION TIMES Mahdi Zhandi1*, Ehsan Seifi-Ghajalo1, Malak Shakeri1, 1 Department of Animal Science, Faculty College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran. Abstract BACKGROUND: Avian sperm is susceptible to lipid peroxidation, compromising their fertility. The semen antioxidant system protects sperm plasma membrane against reactive oxygen species. OBJECTIVE: The study evaluates the effect of glutathione (GSH) addition to semen extender during different equilibration times (ET) on rooster sperm cryopreservation. MATERIALS AND METHODS: Semen samples are weekly collected from 60-week-old broiler breeder roosters. Collected samples were pooled and divided to six equal parts and frozen according to a randomized design (2 × 3 factorial arrangement). Treatments included adding two levels of GSH [0 (GSH-0) or 1 (GSH-1) mM] to semen extender during three ET: 0 (ET-0), 4 (ET-4) or 8 (ET-8) hours. Post-thawed motility and velocity parameters, apoptotic like changes, plasma membrane functionality, and mitochondrial membrane potential (MMP) were evaluated. RESULTS: Post-thawed total motility is improved in the GSH-1 compared to the GSH-0 group (P<0.10). Total motility responded quadratically to increasing levels of ET such that the highest value is recorded at ET-0. Although progressive motility (PM) is not affected by GSH or ET, the highest PM is obtained in the GSH-1×ET-0 group (P<0.05). The VAP and STR is improved in the GSH-1 compared to GSH-0 group; however, VAP decreases quadratically, and STR decreases linearly as ET is advanced (P<0.05). The interactive effect of GSH by ET tends (P<0.08) to affect the wobble coefficient (WOB), such that the highest value is recorded in the GSH-1×ET-0 group. Within both GSH supplemented and control groups, the amplitude of lateral head displacement (ALH) is highest (P<0.05) in the ET-0 group. The percentage of live spermatozoa quadratically decreases and the percentage of dead sperm quadratically increases in response to graded levels of ET (P<0.01). The highest plasma membrane functionality is also noted in the GSH-1×ET-0 group (P<0.05). Mitochondrial membrane potential quadratically decreases in response to increasing levels of ET (P<0.05). CONCLUSION: Generally, GSH supplementation to rooster sperm extender has some beneficial effects on post-thawed sperm motion characteristics, but does not positively interact with ET. Keywords: cryopreservation; glutathione; rooster, semen extender.
CryoLetters 41 (2), 100-105 (2020) HYPOTHERMIC STORAGE OF HUMAN UMBILICAL CORD MESENCHYMAL STEM CELLS AND THEIR HYDROGEL CONSTRUCTS Xiaozhang Zhang1, Gang Zhao1,2* 1 Centre for Biomedical Engineering, Department of Electronic Science & Technology, University of Science and Technology of China, Anhui, Hefei, P.R. China; Abstract BACKGROUND: Human umbilical cord mesenchymal stem cells (hUMSCs) and hUMSCs-hydrogel constructs have great potential in clinical application. However, research about hypothermic storage of hUMSCs and hUMSCs-hydrogel constructs has not been fully investigated. OBJECTIVE: To evaluate the cell survival without and with encapsulation (alginate, alginate with PVA) at 4 ºC and 25 ºC, respectively. MATERIALS AND METHODS: Acridine orange/ethidium bromide (AO/EB) staining and fluorescence staining were used to evaluate cell viability during hypothermic storage. RESULTS: Cells with encapsulation show significantly higher viability than cells without encapsulation at 4 ºC, while there was no obvious difference between viability of cells without and with encapsulation at 25 ºC. In addition, cells encapsulated with alginate and PVA show better viability than cells encapsulated with alginate alone. Moreover, cell viability at 25 ºC was significantly higher than cell viability at 4 ºC. CONCLUSION: hUMSCs aggregates and hUMSCs-hydrogel constructs can survive for 7 days by hypothermic storage at 25 ºC, which facilitates the cell delivery and application of hUMSCs and hUMSCs-hydrogel constructs in clinical medicine. Keywords: human umbilical cord mesenchymal stem cells, alginate, PVA, encapsulation, hypothermic storage.
CryoLetters 41 (2), 106-1147 (2020) EFFECT OF DIFFERENT CONCENTRATIONS OF FRUCTOSE AND GLYCEROL IN TRIS CITRIC ACID EXTENDER ON POST THAW QUALITY AND FERTILITY OF BUFFALO BULL SPERMATOZOA Hussain Ahmed1,2*, Sarwat Jahan2, Mian Muhammad Salman3 1 Department of Zoology, University of Buner, Khyber Pakhtunkhwa (KP), Pakistan. Abstract BACKGROUND: Fructose is considered a vital energy source for metabolic events occurring naturally in the seminal plasma of buffalo spermatozoa. OBJECTIVE: To explore the effect of different concentrations of fructose and glycerol in tris citric acid extender on post thaw quality and in vivo fertility of buffalo spermatozoa. MATERIALS AND METHODS: Semen was collected from three bulls through artificial vagina (42°C). Two ejaculates were collected from each bull per collection day and were evaluated initially for consistency, volume, motility and concentration, followed by dilution in five extenders with supplements (Treatment 1: F0.1,G7 = fructose 0.1% + glycerol 7%; T2: F0.2,G7= fructose 0.2% + glycerol 7%; T3: F0.4,G6.5 = fructose 0.4% + glycerol 6.5%; T4: F0.8,G6 = fructose 0.8%, glycerol 6%; T5: F1.0,G5 = fructose 1% + glycerol 5%). The experiment was replicated four times and the data were assessed with ANOVA. RESULTS: The results showed that percent progressive motility, plasma membrane integrity and supra-vital plasma membrane integrity of spermatozoa was significantly higher (P < 0.05) in extender supplemented with T5 than T1 and T2. Sperm hypo-resistivity, acrosome integrity and DNA integrity were significantly higher in extender supplemented with T5 than T1. Moreover, sperm in vitro quality was significantly higher in T5 than T1 during 30 and 60 min of incubation at 37ºC. Sperm in vivo fertility was significantly higher in extenders supplemented with T5 (57.3%) as compared to T1 (41.3%). CONCLUSION: It is concluded that extender supplemented with T5 improved post thaw semen quality and in vivo fertility of buffalo bull. Keywords: buffalo spermatozoa, glycerol, fructose, plasma membrane integrity, motility, fertility. |
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For Abstracts published from meetings, such as SLTB meetings, go to the
relevant Volume Year of the journal (above). For Full text Free Access Content (from 2000 onwards) go to CryoLetters at Ingenta and look for the blue symbol. |
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