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Mural granulosa cells support to maintain the viability of growing porcine oocytes and its developmental competence after insemination

  • Reproductive Physiology and Disease
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

To enhance the in vitro growth of porcine oocytes, we studied the effect of mural granulosa cells (MGCs) on the viability of oocytes attached to granulosa cells (oocyte-granulosa cell complexes, OGCs) that were obtained from early antral follicles.

Methods and results

When OGCs were cultured with MGCs for 12 days, there were significant improvement (P < 0.05) in the robustness of gap junctional communication between the oocyte and the granulosa cells (82% vs. 59%), the survival rate of oocytes (57% vs. 39%), and the diameter of survived oocytes (118 μm vs. 112 μm). The rate of oocyte release of OGCs cultured with MGCs on the 12th day (1.9%) was significantly lower than that of OGCs cultured without MGCs (26%). Complete meiotic arrest was maintained in the group with MGCs (100%), while partial resumption of spontaneous meiosis was noticed in the absence of MGCs (10–19%). Furthermore, the presence of MGCs increased the oocyte maturation rate after maturation culture in both 12- and 14-day culture groups (P < 0.05, 85–88%) compared to OGCs cultured without MGCs (48–60%). MGCs also significantly improved the blastocyst formation rate (day 7) after ICSI (P < 0.05).

Conclusions

The data of this study thus shows that the presence of MGCs during in vitro oocyte growth plays a crucial role in supporting the developmental competence of growing porcine oocytes attached to the granulosa cells via enhancement of their viability.

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Data and materials availability

The data supporting the findings of the current study are available from the corresponding author upon request.

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Acknowledgments

The authors thank Drs. USK Gamage and A. Takeshita for their helpful comments and M Brahmajosyula, PhD, for editing a draft of this manuscript.

Funding

Part of this work was supported by a grant from the Japan Society for the Promotion of Science (KAKENHI 20K09674 to S.H.) and IVF Japan group.

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Authors and Affiliations

  1. Reproductive Science Laboratory, Graduate School of Medicine, Osaka City University, Osaka, 545-8585, Japan

    Takayuki Yamochi & Shu Hashimoto

  2. IVF Namba Clinic, Osaka, 550-0015, Japan

    Takayuki Yamochi & Shu Hashimoto

  3. HORAC Grand Front Osaka Clinic, Osaka, 530-0011, Japan

    Yoshiharu Morimoto

Authors
  1. Takayuki Yamochi
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  2. Shu Hashimoto
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  3. Yoshiharu Morimoto
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Contributions

TY and SH designed the experiment, interpreted the results, and wrote the manuscript with help from all authors. YM supervised the project.

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Correspondence to Shu Hashimoto.

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Supplementary Information

ESM 1

Supplementary figure Representative images of culturing oocytes. a Intact oocyte on day 10. b Shrunken oocyte on day 6. c Fragmented oocyte on day 6. The region of cell membrane is clearly visible under an interference phase contrast microscope even after granulosa cell proliferation. Bar: 20 μm (PNG 1725 kb).

ESM 2

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Yamochi, T., Hashimoto, S. & Morimoto, Y. Mural granulosa cells support to maintain the viability of growing porcine oocytes and its developmental competence after insemination. J Assist Reprod Genet 38, 2591–2599 (2021). https://doi.org/10.1007/s10815-021-02212-2

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  • DOI: https://doi.org/10.1007/s10815-021-02212-2

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