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Engineering Functional Rat Ovarian Spheroids Using Granulosa and Theca Cells

  • Regenerative Medicine: Original Article
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Abstract

Although menopausal hormone therapy (MHT) is the most effective approach to managing the loss of ovarian activity, serious side effects have been reported. Cell-based therapy is a promising alternative for MHT. This study constructed engineered ovarian cell spheroids and investigated their endocrine function. Theca and granulosa cells were isolated from ovaries of 10-week-old rats. Two types of engineered ovarian cell spheroids were fabricated through forced aggregation in microwells, multilayered spheroids with centralized granulosa aggregates surrounded by an outer layer of theca cells and mixed ovarian spheroids lacking spatial rearrangement. The ovarian cell spheroids were encapsulated into a collagen gel. Non-aggregated ovarian cells served as controls. The endocrine function of the engineered ovarian spheroids was assessed over 30 days. The structure of the spheroids was well maintained during culture. The secretion of 17β-estradiol from both types of engineered ovarian cell spheroids was higher than in the control group and increased continuously in a time-dependent manner. Secretion of 17β-estradiol in the multi-layered ovarian cell spheroids was higher than in the non-layered constructs. Increased secretion of progesterone was detected in the multi-layered ovarian cell spheroids at day 5 of culture and was sustained during the culture period. The initial secretion level of progesterone in the non-layered ovarian cell spheroids was similar to those from the controls and increased significantly from days 21 to 30. An in vitro rat model of engineered ovarian cell spheroids was developed that was capable of secreting sex steroid hormones, indicating that the hormone secreting function of ovaries can be recapitulated ex vivo and potentially adapted for MHT.

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Funding

This study was supported by Jack and Pamela Egan.

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Author notes

  1. Myung Jae Jeon and Young Sik Choi contributed equally to this work.

Authors and Affiliations

  1. Wake Forest Institute for Regenerative Medicine, Wake Forest University School of Medicine, Winston-Salem, NC, 27101, USA

    Myung Jae Jeon, Young Sik Choi, Tracy Criswell, Anthony Atala, James J. Yoo & John D. Jackson

  2. Department of Obstetrics and Gynecology, Seoul National University College of Medicine, 101, Daehak-ro, Jong-gu, Seoul, 110-799, Republic of Korea

    Myung Jae Jeon

  3. Department of Obstetrics and Gynecology, Yonsei University College of Medicine, Seoul, 120-752, Republic of Korea

    Young Sik Choi

  4. Institute of Women’s Life Medical Science, Yonsei University College of Medicine, Seoul, 120-752, Republic of Korea

    Young Sik Choi

  5. Department of Obstetrics and Gynecology, Bundang Jeaseng General Hospital, 20, Seohyeon-ro 180beon-gil, Bundang-gu, Seognam-si, Gyeonggi-do, 13590, Republic of Korea

    Il Dong Kim

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Contributions

Myung Jae Jeon, Young Sik Choi, Anthony Atala, James J. Yoo, and John D. Jackson contributed to study conception and design. Myung Jae Jeon, Young Sik Choi, and Il Dong Kim prepared materials, conducted experiments, and analyzed data. Myung Jae Jeon and Young Sik Choi wrote the first draft of the manuscript. Tracy Criswell, Il Dong Kim, Anthony Atala, James J. Yoo, and John D. Jackson reviewed and commented on previous versions of the manuscript.

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Correspondence to John D. Jackson.

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No humans or human tissues were used in this research. All animal studies were conducted with the approval of the Wake Forest University Health Sciences Animal Care and Use committee. The IACUC protocol number is A14-126.

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Jeon, M.J., Choi, Y.S., Kim, I.D. et al. Engineering Functional Rat Ovarian Spheroids Using Granulosa and Theca Cells. Reprod. Sci. 28, 1697–1708 (2021). https://doi.org/10.1007/s43032-020-00445-7

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