JAK/STAT3 pathway promotes proliferation of ovarian aggregate-derived stem cells in vitro

Maryam Saber; Faezeh Shekari; Seyed-Ahmad Mousavi; Ashraf Moini; Monireh-Sadat Miri; Fereshteh Esfandiari

doi:10.1016/j.yexcr.2023.113689

JAK/STAT3 pathway promotes proliferation of ovarian aggregate-derived stem cells in vitro

Exp Cell Res. 2023 Sep 1;430(1):113689. doi: 10.1016/j.yexcr.2023.113689. Epub 2023 Jun 22.

Authors

Maryam Saber¹, Faezeh Shekari¹, Seyed-Ahmad Mousavi¹, Ashraf Moini², Monireh-Sadat Miri¹, Fereshteh Esfandiari³

Affiliations

¹ Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran.
² Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, ACECR, Tehran, Iran; Department of Gynecology and Obstetrics, Arash Women's Hospital, Tehran University of Medical Sciences, Tehran, Iran; Breast Disease Research Center (BDRC), Tehran University of Medical Sciences, Tehran, Iran.
³ Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran. Electronic address: fereshtehesfandiari@royaninstitute.org.

PMID: 37355151
DOI: 10.1016/j.yexcr.2023.113689

Abstract

Background: The accurate identification and isolation of ovarian stem cells from mammalian ovaries remain a major challenge because of the lack of specific surface markers and suitable in vitro culture systems. Optimized culture conditions for in vitro expansion of ovarian stem cells would allow for identifying requirements of these stem cells for proliferation and differentiation that would pave the way to uncover role of ovarian stem cells in ovarian pathophysiology. Here, we used three-dimensional (3D) aggregate culture system for enrichment of ovarian stem cells and named them aggregate-derived stem cells (ASCs). We hypothesized that mimicking the ovarian microenvironment in vitro by using an aggregate model of the ovary would provide a suitable niche for the isolation of ovarian stem cells from adult mouse and human ovaries and wanted to find out the main cellular pathway governing the proliferation of these stem cells.

Results: We showed that ovarian aggregates take an example from ovary microenvironment in terms of expression of ovarian markers, hormone secretion and supporting the viability of the cells. We found that aggregates-derived stem cells proliferate in vitro as long-term while remained expression of germline markers. These ovarian stem cells differentiated to oocyte like cells in vitro spontaneously. Transplantation of these stem cells in to chemotherapy mouse ovary could restore ovarian structure. RNA-sequencing analysis revealed that interleukin6 is upregulated pathway in ovarian aggregate-derived stem cells. Our data showed that JAK/Stat3 signaling pathway which is activated downstream of IL6 is critical for ovarian stem cells proliferation.

Conclusions: We developed a platform that is highly reproducible for in vitro propagation of ovarian stem cells. Our study provides a primary insight into cellular pathway governing the proliferation of ovarian stem cells.

Keywords: Aggregate; JAK/STAT3 signaling pathway; Ovarian stem cells; Three-dimensional culture system.

Publication types

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

MeSH terms

Adult
Animals
Cell Proliferation
Female
Germ Cells / metabolism
Humans
Mammals / metabolism
Mice
Oocytes* / metabolism
Ovary* / metabolism
STAT3 Transcription Factor / genetics
STAT3 Transcription Factor / metabolism
Stem Cells

Substances

STAT3 protein, human
STAT3 Transcription Factor