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Capacitation IVM improves cumulus function and oocyte quality in minimally stimulated mice

Abstract

Purpose

Oocyte in vitro maturation (IVM) is a patient-friendly reproductive technology but lower success rates than IVF have limited its uptake. Capacitation-IVM (CAPA-IVM) is an innovative new IVM system currently undergoing clinical evaluation. This study aimed to determine temporal effects of the pre-IVM phase of CAPA-IVM on cumulus function and oocyte developmental competence in mildly-stimulated mice.

Methods

Immature cumulus oocyte complexes (COCs) derived from mildly stimulated (23 h PMSG) 28-day-old mice underwent pre-IVM for 0–24 h in medium containing c-type natriuretic peptide (CNP), E2, FSH and insulin, prior to IVM (CAPA-IVM). The effect of pre-IVM duration on cumulus cell function and embryo development post-CAPA-IVM/IVF was assessed.

Results

Day 6 blastocyst rate increased incrementally with increasing pre-IVM duration: 40.6 ± 2.0%, 45.8 ± 1.2%, 52.2 ± 3.5%, 53.3 ± 5.9%, and 59.9 ± 2.5% for 0, 2, 6, 12, and 24 h pre-IVM, respectively (P < 0.01). DNA content/COC, a measure of cumulus cell proliferation, was significantly higher with 24 h pre-IVM group compared to 0, 2, or 6 h pre-IVM (P < 0.001). Pre-IVM for 24 h significantly increased cumulus expansion and mRNA expression of matrix genes Has2 and Tnfaip6 and Areg relative to no pre-IVM control (P < 0.01). Cumulus-oocyte gap-junctional communication (GJC) was maintained throughout 24 h pre-IVM (P < 0.0001), and GJC loss was slowed during the subsequent IVM phase, whilst meiotic resumption was accelerated (P < 0.05). Pre-IVM increased COC ATP and ADP content (P < 0.05), but not AMP, ATP/ADP, and energy charge.

Conclusion

The pre-IVM phase of CAPA-IVM improves the quality of IVM oocytes in a temporally dependent manner and significantly influences cumulus cell function including increased cell proliferation, cumulus expansion, and prolonged cumulus-oocyte GJC.

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Acknowledgments

The authors are grateful for the helpful advice from Dr Sergio Romero (Centro de Fertilidad y Reproduccion Asistida, Lima, Peru), and support from Professor Johan Smitz from the Free University of Brussels, Belgium. The authors thank Sonia Bustamante for technical assistance with LC-MS/MS. Mass spectrometric results were obtained at the Bioanalytical Mass Spectrometry Facility within the Analytical Centre of the University of New South Wales.

Funding

This work was supported by grants (APP1076004, APP1062762, APP1139763) and fellowships (APP1023210, APP1117538) awarded to RBG from the National Health and Medical Research Council of Australia, and from Strategic Funds from the University of New South Wales Sydney. YZ and XL are supported by Overseas Training Funding of Tongji Hospital, China, and Fujian Overseas Study Scholarship China, respectively.

Author information

This study was funded by RBG, and designed by RBG, DR, and YZ. YZ performed all experiments under guidance from DR and MJB and with assistance from XL, DR, and MJB. AEK performed COC DNA analyses and DR performed the LC-MS/MS. YZ and RBG wrote the manuscript which was edited and approved by all authors.

Correspondence to R.B. Gilchrist.

Ethics declarations

All mouse procedures were approved by the UNSW Animal Ethics Committee (ethics number 17/105A) and followed the Australian Code of Practice for the Care and Use of Animals for Scientific Purposes.

Conflict of interest

The authors declare that they have no conflict of interest.

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Zhao, Y., Liao, X., Krysta, A. et al. Capacitation IVM improves cumulus function and oocyte quality in minimally stimulated mice. J Assist Reprod Genet 37, 77–88 (2020). https://doi.org/10.1007/s10815-019-01610-x

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Keywords

  • Oocyte in vitro maturation (IVM)
  • Capacitation-IVM (CAPA-IVM)
  • Pre-maturation (pre-IVM)
  • c-type natriuretic peptide (CNP)
  • Cumulus-oocyte gap junction communication
  • Oocyte quality