Volume 23, Issue 1 (January 2025)
IJRM 2025, 23(1): 33-44 |
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Somayyeh Sadat Tahajjodi1 
, Ehsan Farashahi Yazd1 , Azam Agharahimi2 , Fatemeh Akyash1 , Fatemeh Hajizadeh-Tafti3 , Jalal Golzadeh3 , Reza Aflatoonian4 , Behrouz Aflatoonian *5
, Ehsan Farashahi Yazd1 , Azam Agharahimi2 , Fatemeh Akyash1 , Fatemeh Hajizadeh-Tafti3 , Jalal Golzadeh3 , Reza Aflatoonian4 , Behrouz Aflatoonian *5
1- 1. Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. 2. Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. 3. Department of Reproductive Biology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
2- Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. & Department of Reproductive Biology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
3- Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
4- Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Centre, Royan Institute for Reproductive Biomedicine, Tehran, Iran.
5- 1. Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. 2. Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. 3. Department of Reproductive Biology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ,b.aflatoonian@ssu.ac.ir
2- Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. & Department of Reproductive Biology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
3- Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
4- Department of Endocrinology and Female Infertility, Reproductive Biomedicine Research Centre, Royan Institute for Reproductive Biomedicine, Tehran, Iran.
5- 1. Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. 2. Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. 3. Department of Reproductive Biology, School of Medicine, Shahid Sadoughi University of Medical Sciences, Yazd, Iran. ,
Abstract: (1897 Views)
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.
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.
This article has been extracted from Ph.D. Thesis. (Somayyeh Sadat Tahajjodi)
Keywords: Cumulus cells, Conditioned medium, In vitro gametogenesis (IVG), Human embryonic stem cells, Female germ cells.
Type of Study: Original Article |
Subject:
Stem Cell & Cloning
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