Basic Study
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Aug 26, 2020; 12(8): 857-878
Published online Aug 26, 2020. doi: 10.4252/wjsc.v12.i8.857
Human embryonic stem cell-derived mesenchymal stem cells improved premature ovarian failure
Khadijeh Bahrehbar, Mojtaba Rezazadeh Valojerdi, Fereshteh Esfandiari, Rouhollah Fathi, Seyedeh-Nafiseh Hassani, Hossein Baharvand
Khadijeh Bahrehbar, Fereshteh Esfandiari, Seyedeh-Nafiseh Hassani, Hossein Baharvand, Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, Tehran 1665659911, Iran
Khadijeh Bahrehbar, Hossein Baharvand, Department of Developmental Biology, University of Science and Culture, Tehran 1665659911, Iran
Mojtaba Rezazadeh Valojerdi, Rouhollah Fathi, Department of Embryology, Reproductive Biomedicine Research Center, Royan Institute for Reproductive Biomedicine, Tehran 1665659911, Iran
Mojtaba Rezazadeh Valojerdi, Department of Anatomy, Faculty of Medical Science, Tarbiat Modares University, Tehran 1665659911, Iran
Author contributions: Bahrehbar K and Esfandiari F were involved in collection and/or assembly of the data and manuscript writing; Bahrehbar K, Rezazadeh Valojerdi M, Baharvand H, Fathi R and Hassani SN were involved in data analysis and interpretation; Esfandiari F, Fathi R and Hassani SN were involved in conception and design; Rezazadeh Valojerdi M and Baharvand H were co-corresponding authors and were involved in conducting the experiments, manuscript proof, administrative and financial support.
Supported by Royan Institute, the National Institute for Medical Research Development, No. 963255; and the Ministry of Health and Medical Education, No. 700/147.
Institutional review board statement: This study was reviewed and approved by the Institutional Review Board at Royan Institute.
Institutional animal care and use committee statement: All animal experiments of the study were reviewed and approved by the Institutional Ethical Committee at Royan Institute.
Conflict-of-interest statement: The authors declare that they have no competing interests.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared according to these guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
Corresponding author: Hossein Baharvand, PhD, Full Professor, Department of Stem Cells and Developmental Biology, Royan Institute for Stem Cell Biology and Technology, Banihashem St., Tehran 1665659911, Iran.
Received: February 22, 2020
Peer-review started: February 22, 2020
First decision: April 29, 2020
Revised: June 1, 2020
Accepted: July 18, 2020
Article in press: July 18, 2020
Published online: August 26, 2020

Premature ovarian failure (POF) affects many adult women less than 40 years of age and leads to infertility. According to previous reports, various tissue-specific stem cells can restore ovarian function and folliculogenesis in mice with chemotherapy-induced POF. Human embryonic stem cells (ES) provide an alternative source for mesenchymal stem cells (MSCs) because of their similarities in phenotype and immunomodulatory and anti-inflammatory characteristics. Embryonic stem cell-derived mesenchymal stem cells (ES-MSCs) are attractive candidates for regenerative medicine because of their high proliferation and lack of barriers for harvesting tissue-specific MSCs. However, possible therapeutic effects and underlying mechanisms of transplanted ES-MSCs on cyclophosphamide and busulfan-induced mouse ovarian damage have not been evaluated.


To evaluate ES-MSCs vs bone marrow-derived mesenchymal stem cells (BM-MSCs) in restoring ovarian function in a mouse model of chemotherapy-induced premature ovarian failure.


Female mice received intraperitoneal injections of different doses of cyclophosphamide and busulfan to induce POF. Either human ES-MSCs or BM-MSCs were transplanted into these mice. Ten days after the mice were injected with cyclophosphamide and busulfan and 4 wk after transplantation of the ES-MSCs and/or BM-MSCs, we evaluated body weight, estrous cyclicity, follicle-stimulating hormone and estradiol hormone concentrations and follicle count were used to evaluate the POF model and cell transplantation. Moreover, terminal deoxynucleotidyl transferase mediated 2-deoxyuridine 5-triphosphate nick end labeling, real-time PCR, Western blot analysis and immunohistochemistry and mating was used to evaluate cell transplantation. Enzyme-linked immunosorbent assay was used to analyze vascular endothelial growth factor, insulin-like growth factor 2 and hepatocyte growth factor levels in ES-MSC condition medium in order to investigate the mechanisms that underlie their function.


The human ES-MSCs significantly restored hormone secretion, survival rate and reproductive function in POF mice, which was similar to the results obtained with BM-MSCs. Gene expression analysis and the terminal deoxynucleotidyl transferase mediated 2-deoxyuridine 5-triphosphate nick end labeling assay results indicated that the ES-MSCs and/or BM-MSCs reduced apoptosis in the follicles. Notably, the transplanted mice generated new offspring. The results of different analyses showed increases in antiapoptotic and trophic proteins and genes.


These results suggested that transplantation of human ES-MSCs were similar to BM-MSCs in that they could restore the structure of the injured ovarian tissue and its function in chemotherapy-induced damaged POF mice and rescue fertility. The possible mechanisms of human ES-MSC were related to promotion of follicular development, ovarian secretion, fertility via a paracrine effect and ovarian cell survival.

Keywords: Premature ovarian failure, Human embryonic stem cells, Chemotherapy drugs, Mesenchymal stem cell, Bone marrow, Apoptosis

Core tip: Transplanted human embryonic stem cells are similar to bone marrow-derived mesenchymal stem cells. They can restore injured ovarian tissue structure and function in chemotherapy-induced premature ovarian failure mice and rescue fertility through the paracrine effect and ovarian cell survival.