In vivo oocyte recovery and in vitro embryo production from bovine donors aspirated at different frequencies or following FSH treatment

1999 ◽  
Vol 51 (5) ◽  
pp. 951-961 ◽  
Author(s):  
K.L Goodhand ◽  
R.G Watt ◽  
M.E Staines ◽  
J.S.M Hutchinson ◽  
P.J Broadbent
1997 ◽  
Vol 47 (1) ◽  
pp. 364 ◽  
Author(s):  
K.L. Goodhand ◽  
R.G. Watt ◽  
M.E. Staines ◽  
L.C. Higgins ◽  
D.F. Dolman ◽  
...  

1996 ◽  
Vol 45 (1) ◽  
pp. 355 ◽  
Author(s):  
K.L. Goodhand ◽  
P.J. Broadbent ◽  
J.S.M. Hutchinson ◽  
R.G. Watt ◽  
M.E. Staines ◽  
...  

1997 ◽  
Vol 47 (1) ◽  
pp. 259 ◽  
Author(s):  
L.M.T.E. Lansbergen ◽  
E.H.A.T. Hanenberg ◽  
A.M. van Wagtendonk-de Leeuw

2004 ◽  
Vol 16 (2) ◽  
pp. 259
Author(s):  
J.R. Miles ◽  
C.E. Farin ◽  
K.F. Rodriguez ◽  
J.E. Alexander ◽  
P.W. Farin

The role of the vascular supply in the development of placentas from embryos produced in vitro is poorly understood. The objective of this study was to determine the effects of in vitro embryo production on morphometry of blood vessels within fetal (cotyledonary) and maternal (caruncular) components of the placentome during late gestation. In vivo-produced embryos were recovered from superovulated Holstein cows on Day 7 after estrus. For in vitro embryo production, oocytes were aspirated from the ovaries of Holstein cows, matured in vitro, and then fertilized. Presumptive zygotes with their cumulus cells were transferred into M-199 with 10% estrus cow serum and cultured for 168h post-insemination. Semen from the same Holstein sire was used for the production of in vivo and in vitro embryos. Single blastocysts from each production system were transferred into the uteri of heifers. On Day 222 of gestation, fetuses and placentas were recovered in utero (in vivo, n=12; in vitro, n=12). Placentomes were collected, fixed and sectioned. Fetal and maternal blood vessels were identified within placentome sections using immunocytochemistry for vascular endothelial growth factor (VEGF) protein. A total of 4.8×105μm2 of tissue were examined from each placentome. Stereological methods were used to determine the volume densities of fetal and maternal blood vessels. Data were analyzed by GLM procedures. Fetuses were heavier (P=0.03) in the in vitro group (20.7±1.0kg, LS mean±SEM) compared to the in vivo group (17.3±1.0kg). Placentas were also heavier (P=0.06) for the in vitro group (2.5±0.2kg) compared to the in vivo group (2.0±0.2kg). Placental efficiency, calculated as fetal weight/placental weight, was similar between the two treatment groups (9.0±0.5 and 8.9±0.5 for in vivo and in vitro, respectively). Fetal vascular volume density in placentomes was not different between the two treatment groups (5.4±0.3% and 5.4±0.3% for in vivo and in vitro, respectively). In contrast, maternal vascular volume density was greater (P=0.02) for placentomes in the in vitro group (5.9±0.3%) compared to in vivo controls (4.9±0.3%). In summary, compared to placentomes from embryos produced in vivo, placentomes from embryos produced in vitro had similar volume density of fetal vessels, but had significantly increased volume density of maternal vessels. Supported by the State of North Carolina.


2014 ◽  
Vol 26 (1) ◽  
pp. 162
Author(s):  
H. Tribulo ◽  
J. Carcedo ◽  
R. Tribulo ◽  
J. Menajovsky ◽  
B. Bernal ◽  
...  

An experiment was designed to evaluate in vivo and in vitro embryo production following the use of frozen–thawed conventional or Y-sexed semen from a Brangus bull with known high fertility. For in vivo embryo production, Brangus heifers (n = 12) were superovulated twice in a crossover design and inseminated with sexed or conventional semen. On Day 0, all heifers received an intravaginal progesterone device (DIB 1 g, Syntex S.A., Buenos Aires, Argentina) and 2.5 mg oestradiol benzoate and 50 mg progesterone (Progestar, Syntex S.A.) by intramuscular injection (IM). On Day 4, heifers were superstimulated with 200 mg of NIH-FSH-P1 Folltropin-V (Bioniche Animal Health, Belleville, Ontario, Canada) in twice-daily decreasing doses over 4 days. In the a.m. and p.m. of Day 6, all heifers received PGF2a (Ciclase, Syntex) and DIBs were removed in the p.m.. In the a.m. of Day 8, heifers received 100 μg de Gonadolerin (Gonasyn, Syntex S.A.) and were randomly allocated to receive either one straw of conventional semen (24 × 106 sperm per dose) 12 and 24 h later or two straws of sexed semen (2.4 × 106 sperm per dose) 18 and 24 h after GnRH. Ova/embryos were collected nonsurgically on Day 15 and evaluated following IETS recommendations. Means were compared by t-test. Mean ( ± s.e.m.) number of ova/embryos, fertilized ova, and transferable embryos were 14.8 ± 2.7, 9.4 ± 1.8, and 7.1 ± 1.7 v. 16.8 ± 3.1, 9.9 ± 2.5, and 8.1 ± 2.0 for donors inseminated with conventional or sexed semen, respectively (P > 0.6). For in vitro production, oocytes were obtained from 50 ultrasound-guided follicle aspiration (OPU) sessions that was performed at random stages of the oestrous cycle and without superstimulation in 22 Brangus cows and heifers. Oocytes were classified and matured in TCM-199 medium with NaHCO3 and supplemented with 1% fetal bovine serum. Semen samples from the same bull used for in vivo embryo production were selected using Percoll and capacitated in Fert medium and used at a final concentration of sperm/mL for nonsexed semen and 2 × 106 sperm mL–1 for sexed semen. After 16 h (sexed) or 18 h (conventional) in Fert medium, zygotes were denuded and cultured in SOF supplemented with 0.4% BSA under oil at 37°C, 5% CO2 and saturated humidity for 7 days. The total number of oocytes matured and fertilized was 528 and 318 for conventional and sexed semen, respectively. Means were compared by t-test and proportions by chi-squared test. Mean (± s.e.m.) number of cleaved zygotes and blastocysts produced per OPU session did not differ between conventional (11.0 ± 1.4 and 7.1 ± 1.0) and sexed (8.7 ± 0.8 and 4.9 ± 0.7; P > 0.2) semen. However, the proportion of cleaved zygotes and blastocysts produced were significantly higher (P < 0.05) with conventional semen (61.2%; 329/538 and 39.4%; 212/538) than with sexed semen (54.4%; 173/318 and 30.8%; 98/318), respectively. In conclusion, comparable number of embryos can be obtained in vivo with sexed or conventional semen from a bull with proven high fertility. However, the proportion of blastocysts produced in vitro is likely to be reduced following the use of sexed as compared with conventional semen from the same bull.


2019 ◽  
Vol 31 (1) ◽  
pp. 192
Author(s):  
R. V. Sala ◽  
L. C. Carrenho-Sala ◽  
M. Fosado ◽  
E. Peralta ◽  
D. C. Pereira ◽  
...  

The benefit of superstimulation with exogenous FSH before ovum pickup for in vitro embryo production has been the subject of significant controversy. In addition, there is limited information on different dose regimens. Thus, the objective of the present study was to evaluate the effect of dose of porcine (p)-FSH during superstimulation before ovum pickup (OPU) on in vitro embryo production in pregnant heifers. Pregnant Holstein heifers (n=36) were assigned to a complete 3×3 crossover design. Three treatment groups were evaluated as follows: p-FSH 0mg (FSH0), p-FSH 160mg (FSH160) and p-FSH 300mg (FSH300). Three sessions of OPU were performed on each animal at 48, 62 and 76 days of gestation, with a washout interval between sessions of 14 days. Follicular wave emergence was synchronized by dominant follicle removal. Heifers in the FSH0 group received no further treatment, whereas the remaining groups received a total of 4 injections 12h apart as follows: FSH160 (48.0, 42.7, 37.3 and 32.0mg) or FSH300 (90.0, 80.0, 70.0 and 60.0mg), beginning 36h after dominant follicle removal. Ovum pickup was performed in all heifers 40h after the last p-FSH injection. Heifers were subjected to OPU for oocyte recovery, and number of follicles was determined. Recovered oocytes were processed and in vitro embryo production performed. Differences between treatment groups were evaluated by generalized linear mixed models. Data are presented (Table 1) as mean±standard error of the mean. There was no effect of days in gestation for any of the outcomes evaluated (P&gt;0.05). Follicle numbers at the time of oocyte recovery were different (P&lt;0.01) between groups. Heifers in the FSH300 group had a greater (P&lt;0.05) number of medium, large and total follicles than heifers in the FSH0 group, whereas heifers in the FSH160 were intermediate. Total number of recovered, viable and cleaved oocytes were greater (P&lt;0.01) in FSH300- than in FSH160- and FSH0-treated heifers. Cleavage rate and blastocyst development rate were not different (P&gt;0.10) between groups. The number of grade 1 and 2 blastocysts was greater in FSH300- than in FSH160- and FSH0-treated heifers (P&lt;0.03). In summary, the use of 300mg of p-FSH before OPU in pregnant heifers increases the number of follicles, oocytes and blastocysts produced per heifer with no detrimental effect on oocyte competence. Table 1.Ovum pickup and in vitro embryo production in pregnant heifers treated with different doses of porcine FSH


2016 ◽  
Vol 19 (10) ◽  
pp. 1091-1095
Author(s):  
Camila Louise Ackermann ◽  
Eduardo Trevisol ◽  
Leticia Ferrari Crocomo ◽  
Tatiana da Silva Rascado ◽  
Rodrigo Volpato ◽  
...  

Objectives The present study investigated the effect of contraceptive treatment with deslorelin acetate on in vitro embryo production and oocyte recovery in domestic queens. Methods Twenty-one mature domestic cats were used. Eleven queens (treated group) and one tom were kept in an experimental cattery, and 10 queens were privately owned (control group). When in interestrus or diestrus (day 0) a deslorelin acetate implant (Suprelorin, 4.7 mg/animal) was inserted into the subcutaneous tissue of the interscapular region in all queens in the treated group. After 6 months of treatment, all animals were ovariohysterectomized, and the ovaries were used for in vitro embryo production. Percentage of cleavage was determined 18 h after oocyte insemination and blastocyst formation was assessed on the eighth day of culture. The rate of cumulus-oocyte complexes (COCs) recovery was analyzed by an unpaired t-test. The cleavage and blastocyst rates were expressed as percentages and analyzed by Fisher’s exact test. All analyses were performed using GraphPad Prism v5.0, with P <0.05 set as the level of significance. Results In the treated group, we recovered 8.3 ± 1.15 grade I COCs per queen; the cleavage rate was 60% and the blastocyst rate was 36%. In the control group, we recovered 18.4 ± 3.21 grade I COCs per queen; the cleavage rate was 55.97% and the blastocyst rate was 34%. Forty percent of treated females did not produce any blastocysts. In the treated group, we observed a significant decrease in COC recovery. Although there was no significant difference in cleavage and blastocyst rates between groups, 40% of treated females did not produce any blastocysts. Conclusions Recovery of grade I COCs is negatively affected by deslorelin treatment in domestic cats. Regarding embryo production, new studies are still necessary to evaluate the success of this technique owing to the individual effect caused by deslorelin acetate.


1997 ◽  
Vol 47 (1) ◽  
pp. 287
Author(s):  
P. Davies ◽  
J. Singh ◽  
J. Thundathil ◽  
G. Brogliatti ◽  
D. Bergfelt ◽  
...  

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