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Blastocoele

The Experts below are selected from a list of 225 Experts worldwide ranked by ideXlab platform

George E. Seidel – 1st expert on this subject based on the ideXlab platform

  • 136 Blastocoele COLLAPSE IMPROVES POST-THAW SURVIVAL OF SLOW FROZEN AND VITRIFIED IN VITRO-PRODUCED BOVINE EMBRYOS
    Reproduction Fertility and Development, 2014
    Co-Authors: J. P. Barfield, George E. Seidel

    Abstract:

    Slow-freeze cryopreservation of in vitro-produced bovine embryos often results in poor post-thaw embryo viability. Results with vitrification tend to be better but do not always produce consistent results for a variety of reasons. Experiments with human and equine embryos have demonstrated improved post-thaw survival when Blastocoele fluid is removed before vitrification. We hypothesised that removing Blastocoele fluid before vitrification or slow-freeze cryopreservation would improve post-thaw survival of in vitro-produced bovine embryos. Three replicates of embryos were generated from abattoir-derived ovaries using standard techniques. Seven days post-fertilization, embryos were evaluated for quality, and grade 1 or 2 embryos were allocated to 1 of 4 treatments: slow-freeze control (n = 103); slow-freeze collapsed (n = 106); vitrified control (n = 117); vitrified collapsed (n = 99). Blastocoele fluid was removed by aspiration with a pipette advanced to the centre of the Blastocoele while the embryos were stabilised with a holding pipette. After collapse, embryos were immediately slow frozen in commercial medium [1.5 M ethylene glycol (EG) + 0.1 M sucrose, Bioniche] in 0.25-mL straws or vitrified in 2 steps: 3 min in 1.5 M EG, 30 s in 7 M EG + 0.6 M galactose + 18% Ficoll in open-pulled straws. After thawing or warming, embryos were cultured in chemically defined medium-2 + 5% FCS in 5% CO2, 5% O2, 90% N2 at 38.5°C. Embryos were evaluated for survival based on re-expansion of the Blastocoele after 24 h in culture. Data were analysed by GLIMMIX (SAS Institute Inc., Cary, NC, USA). There were significant replicate effects but there were no significant differences in survival between embryos that were vitrified or slow frozen. Approximately twice as many collapsed embryos survived compared to intact control embryos regardless of whether the embryos were vitrified or slow frozen (collapsed: 50.5 ± 4.1%, control: 26.3 ± 3.5%; P 

  • 136 Blastocoele collapse improves post thaw survival of slow frozen and vitrified in vitro produced bovine embryos
    Reproduction Fertility and Development, 2014
    Co-Authors: J. P. Barfield, George E. Seidel

    Abstract:

    Slow-freeze cryopreservation of in vitro-produced bovine embryos often results in poor post-thaw embryo viability. Results with vitrification tend to be better but do not always produce consistent results for a variety of reasons. Experiments with human and equine embryos have demonstrated improved post-thaw survival when Blastocoele fluid is removed before vitrification. We hypothesised that removing Blastocoele fluid before vitrification or slow-freeze cryopreservation would improve post-thaw survival of in vitro-produced bovine embryos. Three replicates of embryos were generated from abattoir-derived ovaries using standard techniques. Seven days post-fertilization, embryos were evaluated for quality, and grade 1 or 2 embryos were allocated to 1 of 4 treatments: slow-freeze control (n = 103); slow-freeze collapsed (n = 106); vitrified control (n = 117); vitrified collapsed (n = 99). Blastocoele fluid was removed by aspiration with a pipette advanced to the centre of the Blastocoele while the embryos were stabilised with a holding pipette. After collapse, embryos were immediately slow frozen in commercial medium [1.5 M ethylene glycol (EG) + 0.1 M sucrose, Bioniche] in 0.25-mL straws or vitrified in 2 steps: 3 min in 1.5 M EG, 30 s in 7 M EG + 0.6 M galactose + 18% Ficoll in open-pulled straws. After thawing or warming, embryos were cultured in chemically defined medium-2 + 5% FCS in 5% CO2, 5% O2, 90% N2 at 38.5°C. Embryos were evaluated for survival based on re-expansion of the Blastocoele after 24 h in culture. Data were analysed by GLIMMIX (SAS Institute Inc., Cary, NC, USA). There were significant replicate effects but there were no significant differences in survival between embryos that were vitrified or slow frozen. Approximately twice as many collapsed embryos survived compared to intact control embryos regardless of whether the embryos were vitrified or slow frozen (collapsed: 50.5 ± 4.1%, control: 26.3 ± 3.5%; P < 0.001). Advanced blastocysts (fully expanded and hatched blastocysts) survived better than early blastocysts (pre-expansion blastocysts and expanding blastocysts, 45.9% ± 3.9 v. 30.0% ± 4.4, respectively; P < 0.01). The only significant interaction (P < 0.05) was that vitrification was superior to freezing for early blastocysts (39.5 ± 5.9% v. 22.1 ± 5.0% survival) but not advanced blastocysts (44.4 ± 5.3% v. 47.4 ± 5.8% survival). Removal of Blastocoele fluid before cryopreservation of in vitro-produced bovine embryos may be a useful technique for improving post-thaw survival regardless of cryopreservation method or blastocyst stage.

Christopher G. Grupen – 2nd expert on this subject based on the ideXlab platform

  • 59 post-thaw survival of vitrified, in vitro-produced porcine embryos: comparisons of vitrification solutions, solution temperature, and Blastocoele collapse methods.
    Reproduction Fertility and Development, 2015
    Co-Authors: Louise K. Bartolac, Cecilia Sjöblom, Christopher G. Grupen

    Abstract:

    Current post-thaw survival rates of vitrified in vitro-produced porcine blastocysts are much lower than those of other domestic species. The main reason for this is the suboptimal development of in vitro-produced porcine embryos, and the endemically high lipid content of porcine oocytes. Currently, there are several vitrification protocols that have been used to successfully vitrify blastocysts from other species, but many of these have not been used to vitrify porcine embryos. Furthermore, the practice of collapsing the Blastocoele cavity before vitrification is used routinely in human clinics and in other domestic animals; however, there has been little data published regarding the collapse of porcine blastocysts before vitrification. In this study we compared several different vitrification protocols containing different constituents and altered constituent concentrations. We also compared 2 methods of Blastocoele collapse before vitrification. The aim of this study was to determine the optimum conditions for cryopreserving porcine embryos. All experiments were performed on Day 7 in vitro-produced porcine blastocysts. In experiment 1 embryos were vitrified in either a standard solution (17% ethylene glycol + 17% dimethyl sulfoxide + 0.4 M sucrose) or 17% ethylene glycol + 17% propandiol + 0.4 M sucrose. In experiment 2 embryos were vitrified in either the standard solution or 17% ethylene glycol + 17% dimethyl sulfoxide + 0.4 M trehalose. In experiment 3 embryos were vitrified in the standard solution at 38.5°C or at room temperature. In experiment 4 embryos were collapsed before vitrification by micro-pipetting or via 2 sucrose solutions. Survival of embryos was determined by the presence of a Blastocoele cavity 24 h after thaw. Data were subjected to ANOVA and an appropriate post-hoc test when differences were found. The post-thaw survival of embryos vitrified in the standard solution (58%) and the propandiol solution (61%) did not differ. Also, the survival rates of blastocysts vitrified using sucrose (72%) and trehalose (72%) were the same. The survival rate of embryos vitrified in warmed media was significantly lower than embryos vitrified in media at room temperature (54 and 71%, respectively; P 

  • 59 post thaw survival of vitrified in vitro produced porcine embryos comparisons of vitrification solutions solution temperature and Blastocoele collapse methods
    Reproduction Fertility and Development, 2015
    Co-Authors: Louise K. Bartolac, Cecilia Sjöblom, Christopher G. Grupen

    Abstract:

    Current post-thaw survival rates of vitrified in vitro-produced porcine blastocysts are much lower than those of other domestic species. The main reason for this is the suboptimal development of in vitro-produced porcine embryos, and the endemically high lipid content of porcine oocytes. Currently, there are several vitrification protocols that have been used to successfully vitrify blastocysts from other species, but many of these have not been used to vitrify porcine embryos. Furthermore, the practice of collapsing the Blastocoele cavity before vitrification is used routinely in human clinics and in other domestic animals; however, there has been little data published regarding the collapse of porcine blastocysts before vitrification. In this study we compared several different vitrification protocols containing different constituents and altered constituent concentrations. We also compared 2 methods of Blastocoele collapse before vitrification. The aim of this study was to determine the optimum conditions for cryopreserving porcine embryos. All experiments were performed on Day 7 in vitro-produced porcine blastocysts. In experiment 1 embryos were vitrified in either a standard solution (17% ethylene glycol + 17% dimethyl sulfoxide + 0.4 M sucrose) or 17% ethylene glycol + 17% propandiol + 0.4 M sucrose. In experiment 2 embryos were vitrified in either the standard solution or 17% ethylene glycol + 17% dimethyl sulfoxide + 0.4 M trehalose. In experiment 3 embryos were vitrified in the standard solution at 38.5°C or at room temperature. In experiment 4 embryos were collapsed before vitrification by micro-pipetting or via 2 sucrose solutions. Survival of embryos was determined by the presence of a Blastocoele cavity 24 h after thaw. Data were subjected to ANOVA and an appropriate post-hoc test when differences were found. The post-thaw survival of embryos vitrified in the standard solution (58%) and the propandiol solution (61%) did not differ. Also, the survival rates of blastocysts vitrified using sucrose (72%) and trehalose (72%) were the same. The survival rate of embryos vitrified in warmed media was significantly lower than embryos vitrified in media at room temperature (54 and 71%, respectively; P < 0.05). In experiment 4 the post-thaw survival of noncollapsed and embryos collapsed via micro-pipetting was similar (55 and 44%, respectively); however, survival tended to decrease if embryos were collapsed using sucrose before vitrification (25%; P = 0.093). These findings indicate there was little impact on post-thaw survival rates due to the vitrification solutions used. However, the temperature of the solutions appears to influence post-thaw survival. Contrary to findings in other species, collapsing the Blastocoele cavity before vitrification did not improve cryosurvivability; it had no effect or, in the case of sucrose, a negative effect on survival. This may be due to the inherent sensitivity of in vitro-produced porcine embryos to manipulations.

J. P. Barfield – 3rd expert on this subject based on the ideXlab platform

  • 136 Blastocoele COLLAPSE IMPROVES POST-THAW SURVIVAL OF SLOW FROZEN AND VITRIFIED IN VITRO-PRODUCED BOVINE EMBRYOS
    Reproduction Fertility and Development, 2014
    Co-Authors: J. P. Barfield, George E. Seidel

    Abstract:

    Slow-freeze cryopreservation of in vitro-produced bovine embryos often results in poor post-thaw embryo viability. Results with vitrification tend to be better but do not always produce consistent results for a variety of reasons. Experiments with human and equine embryos have demonstrated improved post-thaw survival when Blastocoele fluid is removed before vitrification. We hypothesised that removing Blastocoele fluid before vitrification or slow-freeze cryopreservation would improve post-thaw survival of in vitro-produced bovine embryos. Three replicates of embryos were generated from abattoir-derived ovaries using standard techniques. Seven days post-fertilization, embryos were evaluated for quality, and grade 1 or 2 embryos were allocated to 1 of 4 treatments: slow-freeze control (n = 103); slow-freeze collapsed (n = 106); vitrified control (n = 117); vitrified collapsed (n = 99). Blastocoele fluid was removed by aspiration with a pipette advanced to the centre of the Blastocoele while the embryos were stabilised with a holding pipette. After collapse, embryos were immediately slow frozen in commercial medium [1.5 M ethylene glycol (EG) + 0.1 M sucrose, Bioniche] in 0.25-mL straws or vitrified in 2 steps: 3 min in 1.5 M EG, 30 s in 7 M EG + 0.6 M galactose + 18% Ficoll in open-pulled straws. After thawing or warming, embryos were cultured in chemically defined medium-2 + 5% FCS in 5% CO2, 5% O2, 90% N2 at 38.5°C. Embryos were evaluated for survival based on re-expansion of the Blastocoele after 24 h in culture. Data were analysed by GLIMMIX (SAS Institute Inc., Cary, NC, USA). There were significant replicate effects but there were no significant differences in survival between embryos that were vitrified or slow frozen. Approximately twice as many collapsed embryos survived compared to intact control embryos regardless of whether the embryos were vitrified or slow frozen (collapsed: 50.5 ± 4.1%, control: 26.3 ± 3.5%; P 

  • 136 Blastocoele collapse improves post thaw survival of slow frozen and vitrified in vitro produced bovine embryos
    Reproduction Fertility and Development, 2014
    Co-Authors: J. P. Barfield, George E. Seidel

    Abstract:

    Slow-freeze cryopreservation of in vitro-produced bovine embryos often results in poor post-thaw embryo viability. Results with vitrification tend to be better but do not always produce consistent results for a variety of reasons. Experiments with human and equine embryos have demonstrated improved post-thaw survival when Blastocoele fluid is removed before vitrification. We hypothesised that removing Blastocoele fluid before vitrification or slow-freeze cryopreservation would improve post-thaw survival of in vitro-produced bovine embryos. Three replicates of embryos were generated from abattoir-derived ovaries using standard techniques. Seven days post-fertilization, embryos were evaluated for quality, and grade 1 or 2 embryos were allocated to 1 of 4 treatments: slow-freeze control (n = 103); slow-freeze collapsed (n = 106); vitrified control (n = 117); vitrified collapsed (n = 99). Blastocoele fluid was removed by aspiration with a pipette advanced to the centre of the Blastocoele while the embryos were stabilised with a holding pipette. After collapse, embryos were immediately slow frozen in commercial medium [1.5 M ethylene glycol (EG) + 0.1 M sucrose, Bioniche] in 0.25-mL straws or vitrified in 2 steps: 3 min in 1.5 M EG, 30 s in 7 M EG + 0.6 M galactose + 18% Ficoll in open-pulled straws. After thawing or warming, embryos were cultured in chemically defined medium-2 + 5% FCS in 5% CO2, 5% O2, 90% N2 at 38.5°C. Embryos were evaluated for survival based on re-expansion of the Blastocoele after 24 h in culture. Data were analysed by GLIMMIX (SAS Institute Inc., Cary, NC, USA). There were significant replicate effects but there were no significant differences in survival between embryos that were vitrified or slow frozen. Approximately twice as many collapsed embryos survived compared to intact control embryos regardless of whether the embryos were vitrified or slow frozen (collapsed: 50.5 ± 4.1%, control: 26.3 ± 3.5%; P < 0.001). Advanced blastocysts (fully expanded and hatched blastocysts) survived better than early blastocysts (pre-expansion blastocysts and expanding blastocysts, 45.9% ± 3.9 v. 30.0% ± 4.4, respectively; P < 0.01). The only significant interaction (P < 0.05) was that vitrification was superior to freezing for early blastocysts (39.5 ± 5.9% v. 22.1 ± 5.0% survival) but not advanced blastocysts (44.4 ± 5.3% v. 47.4 ± 5.8% survival). Removal of Blastocoele fluid before cryopreservation of in vitro-produced bovine embryos may be a useful technique for improving post-thaw survival regardless of cryopreservation method or blastocyst stage.