The SLTB 2019 scientific meeting was held on 2-4th October, in the historic Antigua de Fabrica Tobacos building of the Universidad of Seville and was hosted by Professor Ramon Risco and Dr Ariadna Corral. The meeting included a workshop on cryopreservation of cell therapy products (2nd October 2019) run in collaboration with the UK’s Stem Cells User Group and the Andalusian Initiative for Advanced Therapies. This represented an ongoing SLTB strategic objective to engage with other scientific groups interested in low temperature biology. Overall, the conference attracted nine invited keynote speakers and 34 free communications representing a scientifically dense programme dealing with a diverse applications and cryobiological issues in cell preservation of a wide range of cells, tissues and organs.

Alpha-tocopheryl succinate, a major chain-splitting antioxidant, is the most effective form of vitamin E and may be used in the semen extender for cryopreservation of buffalo spermatozoa.

To use different concentrations of alpha-tocopheryl succinate (T1, 0.3 mM, T2, 0.6 mM, and T3, 0.9 mM) and control (0.0 mM) in extender for dose optimization and hence improve the frozen-thawed quality of water buffalo spermatozoa.

Semen samples were collected from three mature buffalo bulls with artificial vagina (42°C) and this study was replicated for five times. Semen was cryopreserved by conventional method which included filling of semen per experimental treatment into 0.5 mL French straws, sealing with polyvinyl alcohol powder and keeping them 5 cm above the liquid nitrogen vapors for 12 min and storing in liquid nitrogen tank. Frozen-thawed semen was also processed for total antioxidant capacity content (TAC) and lipid peroxidation (LPO) level by thiobarbituric acid (TBA). Computer-assisted semen analysis (CASAg the TAC levels and keeping the LPO levels lower as compared to the control. It is suggested that future study should be aimed to explore the influence of these optimal concentrations of alpha-tocopheryl succinate on in vivo fertility of buffalo bull spermatozoa.

The supplementation of alpha-tocopheryl succinate in extender, either at 0.6 (T2) or 0.9 (T3) mM concentrations improves the post thaw quality of water buffalo spermatozoa by sustaining the TAC levels and keeping the LPO levels lower as compared to the control. It is suggested that future study should be aimed to explore the influence of these optimal concentrations of alpha-tocopheryl succinate on in vivo fertility of buffalo bull spermatozoa.

Noninvasive monitoring of cryosurgery is important for performing precise monitoring of the freezing process in situ and evaluating postoperative effects after therapy. One potential approach is to monitor the normal and freeze-thawed tissues through ultrasonic backscattered signal processing.

A noninvasive method for cryosurgery monitoring based on the analysis of microstructural characteristics of in vitro porcine liver tissues at different state including normal and freeze-thawed tissues by estimating the center frequency of scatterers (CFS) using the autoregressive (AR) cepstrum of ultrasonic backscattered signals.

The method is based on the discrete scattering model described in the tissue characterization literature and the observation that most biological tissues are semi-regular scattering lattices. this website A total of ten in vitro porcine liver samples were used and freeze by water bath in the experiments.

Experimental results show that the CFS in porcine liver tissues decreases after pre-frozen and then thawed.

The CFS obtained using this method may be used as a characteristic parameter for tissue characterization in noninvasive monitoring the transition zone between frozen and unfrozen tissues during the surgical therapy, and evaluating postoperative effects.

The CFS obtained using this method may be used as a characteristic parameter for tissue characterization in noninvasive monitoring the transition zone between frozen and unfrozen tissues during the surgical therapy, and evaluating postoperative effects.

Cryopreservation of embryos is of considerable relevance for the implementation of embryo transfer programs and the establishment of embryo banks in several mammalian species.

The present investigation compares two different vitrification systems and two different warming solutions.

Vitrification was performed using Open Pulled Straw (OPS) or CVM RingFibre plug (CVM) devices. Warming was carried out either in a warming solution containing 0.33 M sucrose or in a solution devoid of sucrose.

Differences between vitrification systems were not significant. Warming in sucrose-containing diluent resulted in an expansion rate of 64%, as compared to 86% in a solution devoid of sucrose; reported hatching rates were 45% vs. 9%, respectively (p<0.05). Upon transfer, implantation rates for OPS- and CVM were 50% and 27%, respectively, compared with 55% for freshly collected embryos. The implantation rate after warming was 43% for sucrose-containing and 33% for sucrose-free medium.

a) both vitrification systems are suitable for vitrifying mouse blastocysts; b) warming in sucrose-free diluent yields better embryo survival rates than in diluent containing 0.33 M sucrose.

a) both vitrification systems are suitable for vitrifying mouse blastocysts; b) warming in sucrose-free diluent yields better embryo survival rates than in diluent containing 0.33 M sucrose.

Some antioxidants have been used in semen extenders to reduce adverse effects caused by excessive reactive oxygen species (ROS) production. The study was carried out to assess the effect of quercetin (QC) antioxidant therapy on goat semen submitted to cryopreservation.

To evaluate the effect of quercetin incorporation in different phases of the cryopreservation process of goat spermatozoa.

Five ejaculates from each of four goats (n= 20) were collected and split into four groups Control (G1), without QC; G2, 15 μM of QC added to semen before centrifugation; G3, 15 μM QC added to semen after centrifugation; G4, 15 μM QC added to semen before centrifugation and 15 μM of QC added to semen after centrifugation (total of 30 μM of QC); and cryopreserved. All semen samples were evaluated after thawing for sperm kinetics, plasma membrane integrity, and ROS levels.

Although lower concentrations of ROS were associated with groups that received antioxidant supplementation (P=0.0213), linear and dose dependent (P<0.