Creating a semi-opened micro-cavity ovary through sacrificial microspheres as an in vitro model for discovering the potential effect of ovarian toxic agents

Min Ye; Yiran Shan; Bingchuan Lu; Hao Luo; Binhan Li; Yanmei Zhang; Zixuan Wang; Yuzhi Guo; Liliang Ouyang; Jin Gu; Zhuo Xiong; Ting Zhang

doi:10.1016/j.bioactmat.2023.02.029

Creating a semi-opened micro-cavity ovary through sacrificial microspheres as an in vitro model for discovering the potential effect of ovarian toxic agents

Bioact Mater. 2023 Mar 7:26:216-230. doi: 10.1016/j.bioactmat.2023.02.029. eCollection 2023 Aug.

Authors

Min Ye^{1

2

3}, Yiran Shan⁴, Bingchuan Lu^{1

2

3}, Hao Luo^{1

2

3}, Binhan Li^{1

2

3}, Yanmei Zhang^{1

2

3}, Zixuan Wang^{1

2

3}, Yuzhi Guo^{1

2

3}, Liliang Ouyang^{1

2

3}, Jin Gu⁴, Zhuo Xiong^{1

2

3}, Ting Zhang^{1

2

3}

Affiliations

¹ Biomanufacturing Center, Department of Mechanical Engineering, Tsinghua University, Beijing, 100084, China.
² Biomanufacturing and Rapid Forming Technology Key Laboratory of Beijing, Beijing, 100084, China.
³ Biomanufacturing and Engineering Living Systems, Innovation International Talents Base (111 Base), Beijing, 100084, China.
⁴ MOE Key Laboratory of Bioinformatics, BNRIST Bioinformatics Division, Department of Automation, Tsinghua University, Beijing, 100084, China.

Abstract

The bio-engineered ovary is an essential technology for treating female infertility. Especially the development of relevant in vitro models could be a critical step in a drug study. Herein, we develop a semi-opened culturing system (SOCS) strategy that maintains a 3D structure of follicles during the culture. Based on the SOCS, we further developed micro-cavity ovary (MCO) with mouse follicles by the microsphere-templated technique, where sacrificial gelatin microspheres were mixed with photo-crosslinkable gelatin methacryloyl (GelMA) to engineer a micro-cavity niche for follicle growth. The semi-opened MCO could support the follicle growing to the antral stage, secreting hormones, and ovulating cumulus-oocyte complex out of the MCO without extra manipulation. The MCO-ovulated oocyte exhibits a highly similar transcriptome to the in vivo counterpart (correlation of 0.97) and can be fertilized. Moreover, we found that a high ROS level could affect the cumulus expansion, which may result in anovulation disorder. The damage could be rescued by melatonin, but the end of cumulus expansion was 3h earlier than anticipation, validating that MCO has the potential for investigating ovarian toxic agents in vitro. We provide a novel approach for building an in vitro ovarian model to recapitulate ovarian functions and test chemical toxicity, suggesting it has the potential for clinical research in the future.

Keywords: Bio-engineered ovary; Drug study; Ovarian model; SOCS; Semi-opened MCO.