International Journal of Reproductive BioMedicine

Published date: Mar 10 2025

Journal Title: International Journal of Reproductive BioMedicine

Issue title: International Journal of Reproductive BioMedicine (IJRM): Volume 23, Issue No. 1

Pages: 33 - 44

DOI: 10.18502/ijrm.v23i1.18186

Somayyeh Sadat TahajjodiStem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd

Ehsan Farashahi YazdStem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd

Azam AgharahimiResearch and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd

Fatemeh AkyashStem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd

Fatemeh Hajizadeh-TaftiStem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd

Jalal GolzadehStem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd

Reza AflatoonianDepartment of Endocrinology and Female Infertility, Reproductive Biomedicine Research Centre, Royan Institute for Reproductive Biomedicine, Tehran

Behrouz Aflatoonianb.aflatoonian@ssu.ac.irStem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd

Background: Studies have shown that embryonic stem cells can differentiate into germ cells either spontaneously or directed by the defined factors or by applying a co-culture method or conditioned medium of stromal cells.

Objective: Here, Yazd4 (human embryonic stem cells [hESC] line; 46, XX) was used to evaluate the effect of human cumulus cells conditioned medium (CCCM) on in vitro germ cell production and development.

Materials and Methods: In this experimental study 2 different complete media were applied, Dulbecco’s modified eagle’s medium + 20% fetal bovin serum and embryoid body (EB) medium (KnockOut serum replacement- hESC without basal fibroblast growth factor). Thus, there are 2 control groups for spontaneous differentiation (SD) (SD-EB and SD-DM) and 2 types of CCCM (CCCM-DM and CCCM-EB) as test groups. 40% dilution of the conditioned medium was applied according to previous reports.

Results: Reverse transcription-quantitative polymerase chain reaction and immunofluorescent staining assays revealed that between groups with similar complete medium, CCCM provides a better condition for in vitro germ cell differentiation from hESCs. Moreover, a comparison between the 2 types of complete media showed that EB medium is more supportive than Dulbecco’s modified eagle’s medium +20% fetal bovin serum.

Conclusion: CCCM with EB medium as a complete medium provides a better condition for in vitro germ cell differentiation from hESCs.

Keywords: Cumulus cells, Conditioned medium, In vitro gametogenesis (IVG), Human embryonic stem cells, Female germ cells

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