Offspring production of ovarian organoids derived from spermatogonial stem cells by defined factors with chromatin reorganization

Huacheng Luo; Xiaoyong Li; Geng G Tian; Dali Li; Changliang Hou; Xinbao Ding; Lin Hou; Qifeng Lyu; Yunze Yang; Austin J Cooney; Wenhai Xie; Ji Xiong; Hu Wang; Xiaodong Zhao; Ji Wu

doi:10.1016/j.jare.2021.03.006

Offspring production of ovarian organoids derived from spermatogonial stem cells by defined factors with chromatin reorganization

J Adv Res. 2021 Mar 17:33:81-98. doi: 10.1016/j.jare.2021.03.006. eCollection 2021 Nov.

Authors

Huacheng Luo¹, Xiaoyong Li¹, Geng G Tian¹, Dali Li², Changliang Hou¹, Xinbao Ding¹, Lin Hou¹, Qifeng Lyu³, Yunze Yang¹, Austin J Cooney⁴, Wenhai Xie¹, Ji Xiong¹, Hu Wang¹, Xiaodong Zhao⁵, Ji Wu^{1

6}

Affiliations

¹ Renji Hospital, Key Laboratory for the Genetics of Developmental & Neuropsychiatric Disorders (Ministry of Education), Bio-X Institutes, School of Medicine, Shanghai Jiao Tong University, Shanghai 200032, China.
² Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai 200241, China.
³ Shanghai Ninth People's Hospital Affiliated Shanghai Jiao Tong University Schoolof Medicine, Shanghai Jiao Tong University, Shanghai 200240, China.
⁴ Department of Molecular and Cellular Biology, Baylor College of Medicine, Houston, TX 77030, USA.
⁵ Shanghai Center for Systems Biomedicine, Shanghai Jiao TongUniversity, Shanghai 200240, China.
⁶ Key Laboratory of Fertility Preservation and Maintenance of Ministry of Education, Ningxia Medical University, Yinchuan 750004, China.

Abstract

Introduction: Fate determination of germline stem cells remains poorly understood at the chromatin structure level.

Objectives: Our research hopes to develop successful offspring production of ovarian organoids derived from spermatogonial stem cells (SSCs) by defined factors.

Methods: The offspring production from oocytes transdifferentiated from mouse SSCs with tracking of transplanted SSCs in vivo, single cell whole exome sequencing, and in 3D cell culture reconstitution of the process of oogenesis derived from SSCs. The defined factors were screened with ovarian organoids. We uncovered extensive chromatin reorganization during SSC conversion into induced germline stem cells (iGSCs) using high throughput chromosome conformation.

Results: We demonstrate successful production of offspring from oocytes transdifferentiated from mouse spermatogonial stem cells (SSCs). Furthermore, we demonstrate direct induction of germline stem cells (iGSCs) differentiated into functional oocytes by transduction of H19, Stella, and Zfp57 and inactivation of Plzf in SSCs after screening with ovarian organoids. We uncovered extensive chromatin reorganization during SSC conversion into iGSCs, which was highly similar to female germline stem cells. We observed that although topologically associating domains were stable during SSC conversion, chromatin interactions changed in a striking manner, altering 35% of inactive and active chromosomal compartments throughout the genome.

Conclusion: We demonstrate successful offspring production of ovarian organoids derived from SSCs by defined factors with chromatin reorganization. These findings have important implications in various areas including mammalian gametogenesis, genetic and epigenetic reprogramming, biotechnology, and medicine.

Keywords: 3D cell culture; Chromatin reorganization; Defined factors; Induced germline stem cells; Offspring production; Ovarian organoids.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adult Germline Stem Cells*
Animals
Cell Culture Techniques, Three Dimensional
Chromatin / genetics
Female
Male
Mice
Organoids
Spermatogonia*

Substances

Chromatin