Effect of Pre-IVM Duration with cAMP Modulators on the Production of Cloned Equine Embryos and Foals
The asynchrony of cytoplasmic and nuclear maturation in cumulus–oocyte complexes (COCs) due to prematurely declining concentrations of cyclic adenosine monophosphate (cAMP) has been shown to result in reduced oocyte developmental competence. The objective of this study was to evaluate the effect of...
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2025-07-01
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| author | Jenin V. Cortez Kylie Hardwicke Carlos E. Méndez-Calderón Christopher G. Grupen |
| author_facet | Jenin V. Cortez Kylie Hardwicke Carlos E. Méndez-Calderón Christopher G. Grupen |
| author_sort | Jenin V. Cortez |
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| description | The asynchrony of cytoplasmic and nuclear maturation in cumulus–oocyte complexes (COCs) due to prematurely declining concentrations of cyclic adenosine monophosphate (cAMP) has been shown to result in reduced oocyte developmental competence. The objective of this study was to evaluate the effect of pre-IVM treatment with cAMP modulators for different durations on the developmental potential of equine oocytes used for cloned embryo production. Collected COCs were transferred to cryovials filled with transport medium at 20–22 °C. Within the cryovials, the COCs were either untreated (Control) for 18 h or treated with 50 µM forskolin and 100 µM 3-isobutyl-1-methylxanthine for the first 4 h (Pre-IVM 4 h) or the entire 18 h (Pre-IVM 18 h). Oocytes were then transferred to maturation medium and incubated for a further 22–24 h at 38.5 °C in 5% CO<sub>2</sub> in air. Somatic cell nuclear transfer embryos were then produced using the meiotically mature oocytes and donor cells from six different fibroblast cell lines. The rates of maturation and embryo development did not differ significantly between the groups, though blastocyst formation tended to be inferior in the Pre-IVM 4 h group compared with the Control group (<i>p</i> = 0.06). Of 67 blastocysts produced, 23 were transferred to recipient mares on Day 4 or 5 post-ovulation. Regarding the pregnancy outcomes, no significant differences were found between the groups, and four viable foals were born, each derived from a different donor cell line. The findings expand on those from previous evaluations of this biphasic IVM system, and indicate that the cAMP-modulating treatments exert limited effects under the pre-IVM conditions used here. |
| format | Article |
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| publishDate | 2025-07-01 |
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| spelling | doaj-art-e17b1d30dfa74d788c401c7da60c5f7a2025-07-11T14:35:07ZengMDPI AGAnimals2076-26152025-07-011513196110.3390/ani15131961Effect of Pre-IVM Duration with cAMP Modulators on the Production of Cloned Equine Embryos and FoalsJenin V. Cortez0Kylie Hardwicke1Carlos E. Méndez-Calderón2Christopher G. Grupen3Sydney School of Veterinary Science, Faculty of Science, The University of Sydney, Camden, NSW 2570, AustraliaCatalina Equine Reproduction Centre, North Richmond, NSW 2754, AustraliaCatalina Equine Reproduction Centre, North Richmond, NSW 2754, AustraliaSydney School of Veterinary Science, Faculty of Science, The University of Sydney, Camden, NSW 2570, AustraliaThe asynchrony of cytoplasmic and nuclear maturation in cumulus–oocyte complexes (COCs) due to prematurely declining concentrations of cyclic adenosine monophosphate (cAMP) has been shown to result in reduced oocyte developmental competence. The objective of this study was to evaluate the effect of pre-IVM treatment with cAMP modulators for different durations on the developmental potential of equine oocytes used for cloned embryo production. Collected COCs were transferred to cryovials filled with transport medium at 20–22 °C. Within the cryovials, the COCs were either untreated (Control) for 18 h or treated with 50 µM forskolin and 100 µM 3-isobutyl-1-methylxanthine for the first 4 h (Pre-IVM 4 h) or the entire 18 h (Pre-IVM 18 h). Oocytes were then transferred to maturation medium and incubated for a further 22–24 h at 38.5 °C in 5% CO<sub>2</sub> in air. Somatic cell nuclear transfer embryos were then produced using the meiotically mature oocytes and donor cells from six different fibroblast cell lines. The rates of maturation and embryo development did not differ significantly between the groups, though blastocyst formation tended to be inferior in the Pre-IVM 4 h group compared with the Control group (<i>p</i> = 0.06). Of 67 blastocysts produced, 23 were transferred to recipient mares on Day 4 or 5 post-ovulation. Regarding the pregnancy outcomes, no significant differences were found between the groups, and four viable foals were born, each derived from a different donor cell line. The findings expand on those from previous evaluations of this biphasic IVM system, and indicate that the cAMP-modulating treatments exert limited effects under the pre-IVM conditions used here.https://www.mdpi.com/2076-2615/15/13/1961in vitro maturation (IVM)biphasic IVMsimulated physiological oocyte maturation (SPOM)somatic cell nuclear transfer (SCNT)blastocystembryo transfer |
| spellingShingle | Jenin V. Cortez Kylie Hardwicke Carlos E. Méndez-Calderón Christopher G. Grupen Effect of Pre-IVM Duration with cAMP Modulators on the Production of Cloned Equine Embryos and Foals Animals in vitro maturation (IVM) biphasic IVM simulated physiological oocyte maturation (SPOM) somatic cell nuclear transfer (SCNT) blastocyst embryo transfer |
| title | Effect of Pre-IVM Duration with cAMP Modulators on the Production of Cloned Equine Embryos and Foals |
| title_full | Effect of Pre-IVM Duration with cAMP Modulators on the Production of Cloned Equine Embryos and Foals |
| title_fullStr | Effect of Pre-IVM Duration with cAMP Modulators on the Production of Cloned Equine Embryos and Foals |
| title_full_unstemmed | Effect of Pre-IVM Duration with cAMP Modulators on the Production of Cloned Equine Embryos and Foals |
| title_short | Effect of Pre-IVM Duration with cAMP Modulators on the Production of Cloned Equine Embryos and Foals |
| title_sort | effect of pre ivm duration with camp modulators on the production of cloned equine embryos and foals |
| topic | in vitro maturation (IVM) biphasic IVM simulated physiological oocyte maturation (SPOM) somatic cell nuclear transfer (SCNT) blastocyst embryo transfer |
| url | https://www.mdpi.com/2076-2615/15/13/1961 |
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