Biphasic CAPA-IVM Improves Equine Oocyte Quality and Subsequent Embryo Development Without Inducing Genetic Aberrations

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Publicat a:International Journal of Molecular Sciences vol. 26, no. 12 (2025), p. 5495-5514
Autor principal: Fakhar-I-Adil, Muhammad
Altres autors: Angel-Velez, Daniel, Araftpoor Emin, Amin Qurratul Ain, Hedia, Mohamed, Bühler Marcel, Gevaert Kris, Menten Björn, Van, Soom Ann, Chuva de Sousa Lopes Susana Marina, Stoop, Dominic, De Roo Chloë, Smits Katrien, Heindryckx Björn
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024 7 |a 10.3390/ijms26125495  |2 doi 
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100 1 |a Fakhar-I-Adil, Muhammad  |u Ghent-Fertility and Stem Cell Team (G-FaST), Department for Reproductive Medicine, Department of Human Structure and Repair, Ghent University Hospital, Corneel Heymanslaan 10, 9000 Ghent, Belgium; muhammad.fakhariadil@ugent.be 
245 1 |a Biphasic CAPA-IVM Improves Equine Oocyte Quality and Subsequent Embryo Development Without Inducing Genetic Aberrations 
260 |b MDPI AG  |c 2025 
513 |a Journal Article 
520 3 |a 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. 
653 |a Physiology 
653 |a Ovaries 
653 |a Cytoplasm 
653 |a Sperm 
653 |a Embryos 
653 |a Reproductive technologies 
653 |a Epigenetics 
653 |a Polycystic ovary syndrome 
653 |a Peptides 
653 |a Horses 
653 |a Kinases 
653 |a Proteomics 
653 |a Cell division 
700 1 |a Angel-Velez, Daniel  |u Department of Internal Medicine, Reproduction and Population Medicine, Ghent University, 9820 Merelbeke, Belgium; daniel.angelvelez@ugent.be (D.A.-V.); qurratulain.amin@ugent.be (Q.A.A.); mohamed.hedia@ugent.be (M.H.); ann.vansoom@ugent.be (A.V.S.); katrien.smits@ugent.be (K.S.) 
700 1 |a Araftpoor Emin  |u VIB-UGent Center for Medical Biotechnology, Vlaams Instituut voor Biotechnologie (VIB), 9052 Ghent, Belgium; emin.araftpoor@ugent.be (E.A.); marcel.buhler@ugent.be (M.B.); kris.gevaert@ugent.be (K.G.) 
700 1 |a Amin Qurratul Ain  |u Department of Internal Medicine, Reproduction and Population Medicine, Ghent University, 9820 Merelbeke, Belgium; daniel.angelvelez@ugent.be (D.A.-V.); qurratulain.amin@ugent.be (Q.A.A.); mohamed.hedia@ugent.be (M.H.); ann.vansoom@ugent.be (A.V.S.); katrien.smits@ugent.be (K.S.) 
700 1 |a Hedia, Mohamed  |u Department of Internal Medicine, Reproduction and Population Medicine, Ghent University, 9820 Merelbeke, Belgium; daniel.angelvelez@ugent.be (D.A.-V.); qurratulain.amin@ugent.be (Q.A.A.); mohamed.hedia@ugent.be (M.H.); ann.vansoom@ugent.be (A.V.S.); katrien.smits@ugent.be (K.S.) 
700 1 |a Bühler Marcel  |u VIB-UGent Center for Medical Biotechnology, Vlaams Instituut voor Biotechnologie (VIB), 9052 Ghent, Belgium; emin.araftpoor@ugent.be (E.A.); marcel.buhler@ugent.be (M.B.); kris.gevaert@ugent.be (K.G.) 
700 1 |a Gevaert Kris  |u VIB-UGent Center for Medical Biotechnology, Vlaams Instituut voor Biotechnologie (VIB), 9052 Ghent, Belgium; emin.araftpoor@ugent.be (E.A.); marcel.buhler@ugent.be (M.B.); kris.gevaert@ugent.be (K.G.) 
700 1 |a Menten Björn  |u Center for Medical Genetics, Ghent University Hospital, 9000 Ghent, Belgium; bjorn.menten@ugent.be 
700 1 |a Van, Soom Ann  |u Department of Internal Medicine, Reproduction and Population Medicine, Ghent University, 9820 Merelbeke, Belgium; daniel.angelvelez@ugent.be (D.A.-V.); qurratulain.amin@ugent.be (Q.A.A.); mohamed.hedia@ugent.be (M.H.); ann.vansoom@ugent.be (A.V.S.); katrien.smits@ugent.be (K.S.) 
700 1 |a Chuva de Sousa Lopes Susana Marina  |u Department of Anatomy and Embryology, Leiden University Medical Center, 2333 ZA Leiden, The Netherlands; s.m.chuva_de_sousa_lopes@lumc.nl 
700 1 |a Stoop, Dominic  |u Department for Reproductive Medicine, Women’s Clinic, Ghent University Hospital, Corneel Heymanslaan 10, 9000 Ghent, Belgium; dominic.stoop@uzgent.be (D.S.); chloe.deroo@uzgent.be (C.D.R.) 
700 1 |a De Roo Chloë  |u Department for Reproductive Medicine, Women’s Clinic, Ghent University Hospital, Corneel Heymanslaan 10, 9000 Ghent, Belgium; dominic.stoop@uzgent.be (D.S.); chloe.deroo@uzgent.be (C.D.R.) 
700 1 |a Smits Katrien  |u Department of Internal Medicine, Reproduction and Population Medicine, Ghent University, 9820 Merelbeke, Belgium; daniel.angelvelez@ugent.be (D.A.-V.); qurratulain.amin@ugent.be (Q.A.A.); mohamed.hedia@ugent.be (M.H.); ann.vansoom@ugent.be (A.V.S.); katrien.smits@ugent.be (K.S.) 
700 1 |a Heindryckx Björn  |u Ghent-Fertility and Stem Cell Team (G-FaST), Department for Reproductive Medicine, Department of Human Structure and Repair, Ghent University Hospital, Corneel Heymanslaan 10, 9000 Ghent, Belgium; muhammad.fakhariadil@ugent.be 
773 0 |t International Journal of Molecular Sciences  |g vol. 26, no. 12 (2025), p. 5495-5514 
786 0 |d ProQuest  |t Health & Medical Collection 
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