Production and Culture of the Bovine Embryo

Tríbulo, Paula; Rivera, Rocío Melissa; Ortega, M. Sofía; Jannaman, Elizabeth; Hansen, Peter J.

doi:10.1007/978-1-4939-9566-0_8

Cited by 55 publications

(44 citation statements)

References 12 publications

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“…Embryos are produced in vivo by treating cows with follicle-stimulating hormone ( FSH ) to induce multiple ovulations followed by insemination at estrus or the anticipated time of ovulation, and recovery of embryos by flushing the uterus at day 7 after estrus ( Mikkola et al, 2019 ). Embryos are produced in vitro by aspiration of oocytes from follicles (usually by ultrasound-guided transvaginal aspiration but sometimes by harvesting oocytes from ovaries ex situ), inducing nuclear and cytoplasmic maturation of the oocytes in culture, incubating the matured oocytes with capacitated sperm to achieve fertilization, and then culturing the resultant zygotes to the morula or blastocyst stage for transfer ( Tríbulo et al, 2019 ).…”

Section: Causes Of Pregnancy Loss Associated With Etmentioning

confidence: 99%

The incompletely fulfilled promise of embryo transfer in cattle—why aren’t pregnancy rates greater and what can we do about it?

Hansen

2020

Journal of Animal Science

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Typically, bovine embryos are transferred into recipient females about day 7 after estrus or anticipated ovulation, when the embryo has reached the blastocyst stage of development. All the biological and technical causes for failure of a female to produce a blastocyst 7 d after natural or artificial insemination (AI) are avoided when a blastocyst-stage embryo is transferred into the female. It is reasonable to expect, therefore, that pregnancy success would be higher for embryo transfer (ET) recipients than for inseminated females. This expectation is not usually met unless the recipient is exposed to heat stress or is classified as a repeat-breeder female. Rather, pregnancy success is generally similar for ET and AI. The implication is that either one or more of the technical aspects of ET have not yet been optimized or that underlying female fertility that causes an embryo to die before day 7 also causes it to die later in pregnancy. Improvements in pregnancy success after ET will depend upon making a better embryo, improving uterine receptivity, and forging new tools for production and transfer of embryos. Key to accelerating progress in improving pregnancy rates will be the identification of phenotypes or phenomes that allow the prediction of embryo competence for survival and maternal capacity to support embryonic development.

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Section: Causes Of Pregnancy Loss Associated With Etmentioning

confidence: 99%

The incompletely fulfilled promise of embryo transfer in cattle—why aren’t pregnancy rates greater and what can we do about it?

Hansen

2020

Journal of Animal Science

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“…Semen was first centrifuged at 1000 × g for 10 min to separate live sperm from dead sperm and to remove the diluent and seminal plasma. Then the resultant pellet at the bottom of the tube was placed in 10 mL of warmed HEPES-TALP (Tríbulo et al 2019) in a 15-mL conical tube and a second centrifugation at 200 × g for 5 min was performed to wash the sperm. Sperm were resuspended in IVF-TALP medium (Tríbulo et al 2019) previously equilibrated in an incubator in a humidified atmosphere of 5% CO 2 in air at 38.5 °C and the sperm concentration adjusted to 16 × 10 6 sperm/mL using a hemocytometer so that final concentration in the fertilization well was 1 × 10 6 /mL.…”

Section: Embryo Productionmentioning

confidence: 99%

“…For fertilization, matured COC were rinsed 3 times with warmed HEPES-TALP and up to 300 matured COC were transferred to a fertilization dish containing 1.7 mL of IVF-TALP, 120 µl of sperm and 80 µl of PHE solution (0.5 mM penicillamine, 0.25 mM hypotaurine, 0.25 µM epinephrine), prepared as described elsewhere (Tríbulo et al 2019). Gametes were co-incubated in a humidified atmosphere of 5% CO 2 in air at 38.5° for 20-22 h.…”

Section: Embryo Productionmentioning

confidence: 99%

“…Cumulus cells were removed from the putative zygotes by agitation for 5 min using a vortex mixer. Putative zygotes were transferred using a wiretrol pipette in groups of 30 into 45-µl drops of Synthetic Oviduct Fluid-Bovine Embryo 2 (SOF-BE2) (Tríbulo et al 2019) covered with mineral. Embryos were cultured for 7 days in a humidified atmosphere of 5% CO 2 , 5% O 2 , 90% N 2 at 38.5 °C in an EVE model benchtop incubator (WTA, Cravinhos, SP, Brazil).…”

Section: Embryo Productionmentioning

confidence: 99%

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Determinants of survival of the bovine blastocyst to cryopreservation stress: treatment with colony stimulating factor 2 during the morula-to-blastocyst transition and embryo sex

et al. 2020

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Background: Colony-stimulating factor 2 (CSF2) is an important maternal regulator of embryonic development. Earlier research indicates that CSF2 can regulate genes involved in cellular stress responses and block apoptosis. Here, we tested whether addition of 10 ng/mL CSF2 at day 5 of development would increase the survival of blastocysts harvested at day 7 and subjected to vitrification. Additional objectives were to determine whether embryo sex affected survival or whether effects of CSF2 interacted with sex. Results: Survival after vitrification was measured as the percent of warmed blastocysts that re-established a blastocoele after culture and that underwent hatching from the zona pellucida. In the first experiment, blastocysts were vitrified, warmed, cultured for 24 h, and DNA embryo sexing performed by PCR. There was no effect of CSF2, sex, or the interaction on the percent of blastocysts that re-expanded or that were hatching or hatched. In the second experiment, vitrified blastocysts were warmed and cultured for 24, 48, and 72 h. Treatment with CSF2 increased (P = 0.021) the percent of blastocysts that re-expanded as compared to the vehicle group (overall, 77.8 ± 4.7% vs 73.3 ± 4.7%). Percent re-expansion was highest at 24 h and declined slightly thereafter (P = 0.024). Although the interaction was not significant, the effect of CSF2 was greater at 48 and 72 h than at 24 h because CSF2 reduced the incidence of embryos collapsing after re-expansion. Furthermore, the proportion of re-expanded blastocysts at 24 h that experienced blastocoel collapse by 72 h was lower (P = 0.053) for CSF2 (3.6%; 7/195) than for vehicle (8.2%; 16/195). The percent of warmed blastocysts that were hatching or hatched increased with time (P < 0.0001) but there was no effect of CSF2 or the interaction with time on hatching. Conclusion: Treatment with CSF2 from day 5 to 7 of development did not cause a significant effect on the percent of blastocysts that re-established the blastocoele after 24 h of culture but CSF2 reduced the collapse of the blastocoele that occurred for a portion of the embryos that had experienced re-expansion at 24 h. Thus, CSF2 can provide protection to a proportion of blastocysts from cryodamage caused by vitrification. Further work is needed to evaluate whether CSF2 increases survival of vitrified blastocysts after embryo transfer.

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“…Lesson from the observation and investigation of normal physiological systems, especially the female reproductive system, has resulted in the formulation of media to improve in vitro culture conditions and suit the embryo's requirements [1]. Based on the literature, it is possible to have a rough estimate of the performance of each media type regarding pre-and post-implantation competence [2].…”

Section: Introductionmentioning

confidence: 99%

The Impact of Two Embryo Culture Media, SOF and Commercial BO, on Pre- and Post-Implantation Development of Cloned Sannen Goat Embryos

Hajian

Jafarpour

Aghamiri

et al. 2020

Preprint

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Background: The ingredients of embryo culture media developed by different companies are disclosed. Thus, it is impossible to determine which ingredients might be responsible for differences in pre-and post-implantation embryo development. To address this gap, we performed an experiment to compare two embryo culture media, namely, SOF and commercial BO, on pre- and post-implantation development of cloned Sannen goat embryos. Cumulus oocyte complexes derived from slaughterhouse ovaries were used for in vitro embryo production . In vitro development of IVF, parthenogenetic and SCNT embryos were assessed in both BO and SOF media. The expression of 16 genes, including AKT , OCT4 , SOX2 , BMPR1 , FGFR4 , CDC25 , CDX2 , GCN5 , PCAF , FOXD3 , SMAD5 , FZD , LIFR1 , CTNNB , ERK1 , and IFNT , belonging to 7 important pathways, i.e. pluripotency, FGF, TGFβ, cell cycle and proliferation, histone transferase, trophectoderm, and WNT, were examined in the goat SCNT and IVF blastocysts from both BO and SOF media. Results: The blastocyst rate in BO medium was significantly higher than that of the SOF medium in SCNT embryos ( P < 0.05). All of the genes examined showed increased expression levels in SCNT embryos compared to IVF embryos. In the IVF group, OCT4 , BMPR1 , and GCN5 showed significantly higher expression in the SOF medium compared to the BO medium. In this group, AKT , FGFR4 , SOX2 showed significantly lower expression in the SOF medium compared to the BO medium. In the SCNT group, FGFR4 , GCN5 , FZD , CTNNB , BMPR1 , and FGFR4 showed significantly higher expression in SOF medium compared to BO medium. In vivo development did not differ significantly between the two groups. Conclusions: Based on these results, we concluded that the limited information available on the allocations of ICM and TE cells in SCNT embryos and embryo-specific gene expression may be the major drawback IVC medium and an impediment to successful animal cloning.

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Production and Culture of the Bovine Embryo

Cited by 55 publications

References 12 publications

The incompletely fulfilled promise of embryo transfer in cattle—why aren’t pregnancy rates greater and what can we do about it?

The incompletely fulfilled promise of embryo transfer in cattle—why aren’t pregnancy rates greater and what can we do about it?

Determinants of survival of the bovine blastocyst to cryopreservation stress: treatment with colony stimulating factor 2 during the morula-to-blastocyst transition and embryo sex

The Impact of Two Embryo Culture Media, SOF and Commercial BO, on Pre- and Post-Implantation Development of Cloned Sannen Goat Embryos

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