Tahajjodi Somayyeh Sadat, Yazd Ehsan Farashahi, Agharahimi Azam, Akyash Fatemeh, Hajizadeh-Tafti Fatemeh, Golzadeh Jalal, Aflatoonian Reza, Aflatoonian Behrouz
Stem Cell Biology Research Center, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Research and Clinical Center for Infertility, Yazd Reproductive Sciences Institute, Shahid Sadoughi University of Medical Sciences, Yazd, Iran.
Int J Reprod Biomed. 2025 Mar 21;23(1):33-44. doi: 10.18502/ijrm.v23i1.18186. eCollection 2025 Jan.
Studies have shown thatembryonic 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.
Here, Yazd4 (human embryonic stem cells [hESC] line; 46, XX) was used to evaluate the effect of human cumulus cells conditioned medium (CCCM) on germ cell production and development.
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.
Reverse transcription-quantitative polymerase chain reaction and immunofluorescent staining assays revealed that between groups with similar complete medium, CCCM provides a better condition for 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.
CCCM with EB medium as a complete medium provides a better condition for germ cell differentiation from hESCs.
研究表明,胚胎干细胞可以自发分化为生殖细胞,也可以由特定因子引导,或者通过共培养方法或应用基质细胞的条件培养基来实现。
本研究使用Yazd4(人胚胎干细胞系;46, XX)来评估人卵丘细胞条件培养基(CCCM)对生殖细胞产生和发育的影响。
在本实验研究中,使用了2种不同的完全培养基,即杜氏改良 Eagle 培养基 + 20% 胎牛血清和胚状体(EB)培养基(无基础成纤维细胞生长因子的 KnockOut 血清替代物 - hESC)。因此,有2个自发分化(SD)对照组(SD-EB 和 SD-DM)以及2种类型的CCCM(CCCM-DM 和 CCCM-EB)作为试验组。根据先前的报道,对条件培养基进行了40% 的稀释。
逆转录 - 定量聚合酶链反应和免疫荧光染色分析表明,在使用相似完全培养基的组之间,CCCM为从人胚胎干细胞分化出生殖细胞提供了更好的条件。此外,对2种完全培养基的比较显示,EB培养基比杜氏改良 Eagle 培养基 + 20% 胎牛血清更具支持性。
以EB培养基作为完全培养基的CCCM为从人胚胎干细胞分化出生殖细胞提供了更好的条件。