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Biphasic CAPA-IVM improves equine oocyte quality and subsequent embryo development without inducing genetic aberrations

Muhammad Fakhar I Adil (UGent) , Daniel Angel Velez (UGent) , Emin Araftpoor (UGent) , Qurratul Ain Amin (UGent) , Mohamed Hedia (UGent) , Marcel Bühler (UGent) , Kris Gevaert (UGent) , Björn Menten (UGent) , Ann Van Soom (UGent) , Susana Marina Chuva de Sousa Lopes (UGent) , et al.
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Abstract
In vitro maturation (IVM) of oocytes retrieved from ovum pick-up (OPU) or ovarian tissue (OT) is a standard approach for patients with specific conditions where prior hormonal stimulation is contraindicated. However, the developmental competence of oocytes matured in vitro is still inferior to that of oocytes matured in vivo. Capacitation IVM (CAPA-IVM) includes an extra step of pre-maturation culture (PMC) with c-type natriuretic peptide (CNP) as a meiotic arrestor to better synchronize cytoplasmic and nuclear maturity in oocytes by allowing the cytoplasm additional time to acquire essential components critical for optimal competency. This study aims to evaluate the effect of CAPA-IVM on equine oocyte quality and developmental competence. Immature cumulus–oocyte complexes (COCs) were retrieved from slaughterhouse ovaries and matured in vitro either in CAPA-IVM (short 6 h, long 24 h pre-maturation) or standard IVM. Mature oocytes from each group were analyzed for calcium-releasing potential (n = 52) and single-oocyte proteomics (n = 44), and embryo development (n = 229) was assessed after fertilization with piezo-drilled intracytoplasmic sperm injection (ICSI). Genetic analysis of developed blastocysts (n = 41) was performed to detect chromosomal aberrations. Our findings demonstrate that CAPA-IVM of equine COCs yields significantly higher maturation rates than controls. Moreover, short CAPA-IVM with six hours pre-maturation culture showed substantially higher embryo development potential than the control group (20/69 vs. 9/63, respectively). Genetic analysis revealed a high euploidy rate in equine blastocysts regardless of the maturation conditions. Live calcium imaging of the fertilized oocytes demonstrated that the majority of oocytes displayed non-continuous calcium oscillation patterns, irrespective of maturation conditions. Single-oocyte proteomics reveals a comparable proteomic landscape between mature oocytes subjected to short CAPA-IVM and standard IVM. However, we identified four enriched gene sets with positive enrichment scores after short CAPA-IVM, related to cytoskeleton regulation, ribosomal function, and cytosolic components. Our findings indicate that CAPA-IVM holds the potential to improve oocyte quality and competence in horses. However, further fine-tuning of culture conditions would benefit the effective use of these IVM systems. Moreover, given that the mare serves as an excellent model for human reproduction, the molecular trends identified in this study could provide valuable insights for advancing human artificial reproductive technologies.
Keywords
CAPA-IVM, equine embryos, in vitro maturation, oocyte proteomics, equine euploidy, IN-VITRO MATURATION, INTRACYTOPLASMIC SPERM INJECTION, INTRACELLULAR CALCIUM OSCILLATIONS, PHOSPHODIESTERASE TYPE-3 INHIBITOR, MEIOTIC ARREST, BLASTOCYST DEVELOPMENT, NATRIURETIC PEPTIDE, FOLLICULAR DEVELOPMENT, CYCLIC-GMP, COMPETENCE

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MLA
Fakhar I Adil, Muhammad, et al. “Biphasic CAPA-IVM Improves Equine Oocyte Quality and Subsequent Embryo Development without Inducing Genetic Aberrations.” INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, vol. 26, no. 12, 2025, doi:10.3390/ijms26125495.
APA
Fakhar I Adil, M., Angel Velez, D., Araftpoor, E., Amin, Q. A., Hedia, M., Bühler, M., … Heindryckx, B. (2025). Biphasic CAPA-IVM improves equine oocyte quality and subsequent embryo development without inducing genetic aberrations. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 26(12). https://doi.org/10.3390/ijms26125495
Chicago author-date
Fakhar I Adil, Muhammad, Daniel Angel Velez, Emin Araftpoor, Qurratul Ain Amin, Mohamed Hedia, Marcel Bühler, Kris Gevaert, et al. 2025. “Biphasic CAPA-IVM Improves Equine Oocyte Quality and Subsequent Embryo Development without Inducing Genetic Aberrations.” INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 26 (12). https://doi.org/10.3390/ijms26125495.
Chicago author-date (all authors)
Fakhar I Adil, Muhammad, Daniel Angel Velez, Emin Araftpoor, Qurratul Ain Amin, Mohamed Hedia, Marcel Bühler, Kris Gevaert, Björn Menten, Ann Van Soom, Susana Marina Chuva de Sousa Lopes, Dominic Stoop, Chloë De Roo, Katrien Smits, and Björn Heindryckx. 2025. “Biphasic CAPA-IVM Improves Equine Oocyte Quality and Subsequent Embryo Development without Inducing Genetic Aberrations.” INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES 26 (12). doi:10.3390/ijms26125495.
Vancouver
1.
Fakhar I Adil M, Angel Velez D, Araftpoor E, Amin QA, Hedia M, Bühler M, et al. Biphasic CAPA-IVM improves equine oocyte quality and subsequent embryo development without inducing genetic aberrations. INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES. 2025;26(12).
IEEE
[1]
M. Fakhar I Adil et al., “Biphasic CAPA-IVM improves equine oocyte quality and subsequent embryo development without inducing genetic aberrations,” INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, vol. 26, no. 12, 2025.
@article{01JXCYGQXM14A3YJQ5QC6PBVHV,
  abstract     = {{In vitro maturation (IVM) of oocytes retrieved from ovum pick-up (OPU) or ovarian tissue (OT) is a standard approach for patients with specific conditions where prior hormonal stimulation is contraindicated. However, the developmental competence of oocytes matured in vitro is still inferior to that of oocytes matured in vivo. Capacitation IVM (CAPA-IVM) includes an extra step of pre-maturation culture (PMC) with c-type natriuretic peptide (CNP) as a meiotic arrestor to better synchronize cytoplasmic and nuclear maturity in oocytes by allowing the cytoplasm additional time to acquire essential components critical for optimal competency. This study aims to evaluate the effect of CAPA-IVM on equine oocyte quality and developmental competence. Immature cumulus–oocyte complexes (COCs) were retrieved from slaughterhouse ovaries and matured in vitro either in CAPA-IVM (short 6 h, long 24 h pre-maturation) or standard IVM. Mature oocytes from each group were analyzed for calcium-releasing potential (n = 52) and single-oocyte proteomics (n = 44), and embryo development (n = 229) was assessed after fertilization with piezo-drilled intracytoplasmic sperm injection (ICSI). Genetic analysis of developed blastocysts (n = 41) was performed to detect chromosomal aberrations. Our findings demonstrate that CAPA-IVM of equine COCs yields significantly higher maturation rates than controls. Moreover, short CAPA-IVM with six hours pre-maturation culture showed substantially higher embryo development potential than the control group (20/69 vs. 9/63, respectively). Genetic analysis revealed a high euploidy rate in equine blastocysts regardless of the maturation conditions. Live calcium imaging of the fertilized oocytes demonstrated that the majority of oocytes displayed non-continuous calcium oscillation patterns, irrespective of maturation conditions. Single-oocyte proteomics reveals a comparable proteomic landscape between mature oocytes subjected to short CAPA-IVM and standard IVM. However, we identified four enriched gene sets with positive enrichment scores after short CAPA-IVM, related to cytoskeleton regulation, ribosomal function, and cytosolic components. Our findings indicate that CAPA-IVM holds the potential to improve oocyte quality and competence in horses. However, further fine-tuning of culture conditions would benefit the effective use of these IVM systems. Moreover, given that the mare serves as an excellent model for human reproduction, the molecular trends identified in this study could provide valuable insights for advancing human artificial reproductive technologies.}},
  articleno    = {{5495}},
  author       = {{Fakhar I Adil, Muhammad and Angel Velez, Daniel and Araftpoor, Emin and Amin, Qurratul Ain and Hedia, Mohamed and Bühler, Marcel and Gevaert, Kris and Menten, Björn and Van Soom, Ann and Chuva de Sousa Lopes, Susana Marina and Stoop, Dominic and De Roo, Chloë and Smits, Katrien and Heindryckx, Björn}},
  issn         = {{1661-6596}},
  journal      = {{INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES}},
  keywords     = {{CAPA-IVM,equine embryos,in vitro maturation,oocyte proteomics,equine euploidy,IN-VITRO MATURATION,INTRACYTOPLASMIC SPERM INJECTION,INTRACELLULAR CALCIUM OSCILLATIONS,PHOSPHODIESTERASE TYPE-3 INHIBITOR,MEIOTIC ARREST,BLASTOCYST DEVELOPMENT,NATRIURETIC PEPTIDE,FOLLICULAR DEVELOPMENT,CYCLIC-GMP,COMPETENCE}},
  language     = {{eng}},
  number       = {{12}},
  pages        = {{19}},
  title        = {{Biphasic CAPA-IVM improves equine oocyte quality and subsequent embryo development without inducing genetic aberrations}},
  url          = {{http://doi.org/10.3390/ijms26125495}},
  volume       = {{26}},
  year         = {{2025}},
}

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