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卵丘细胞条件培养基促进人胚胎干细胞向生殖细胞分化：一项实验研究。

Cumulus cells conditioned medium facilitates germ cell differentiation from human embryonic stem cells: An experimental study.

作者信息

Tahajjodi Somayyeh Sadat, Yazd Ehsan Farashahi, Agharahimi Azam, Akyash Fatemeh, Hajizadeh-Tafti Fatemeh, Golzadeh Jalal, Aflatoonian Reza, Aflatoonian Behrouz

机构信息

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.

出版信息

Int J Reprod Biomed. 2025 Mar 21;23(1):33-44. doi: 10.18502/ijrm.v23i1.18186. eCollection 2025 Jan.

DOI:10.18502/ijrm.v23i1.18186
PMID:40190455
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11966215/
Abstract

BACKGROUND

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.

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 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 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 germ cell differentiation from hESCs.

摘要

背景

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。

目的

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。

材料与方法

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。

结果

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。

结论

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53d2/11966215/150f53b412f3/ijrb-23-33-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53d2/11966215/9a0949845c07/ijrb-23-33-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53d2/11966215/a392a0c7e0ac/ijrb-23-33-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53d2/11966215/12701d043ff7/ijrb-23-33-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53d2/11966215/9e02316bab69/ijrb-23-33-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53d2/11966215/8644a4033404/ijrb-23-33-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53d2/11966215/150f53b412f3/ijrb-23-33-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53d2/11966215/9a0949845c07/ijrb-23-33-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53d2/11966215/a392a0c7e0ac/ijrb-23-33-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53d2/11966215/12701d043ff7/ijrb-23-33-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53d2/11966215/9e02316bab69/ijrb-23-33-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53d2/11966215/8644a4033404/ijrb-23-33-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/53d2/11966215/150f53b412f3/ijrb-23-33-g006.jpg

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Xeno-free generation of new Yazd human embryonic stem cell lines (Yazd4-7) as a prior stage toward good manufacturing practice of clinical-grade raw materials from discarded embryos: A lab resources report.无动物源生成新的亚兹德人类胚胎干细胞系（Yazd4 - 7）：迈向从废弃胚胎中获取临床级原材料良好生产规范的前期阶段——一份实验室资源报告
Int J Reprod Biomed. 2023 Sep 20;21(8):619-628. doi: 10.18502/ijrm.v21i8.14017. eCollection 2023 Aug.
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Established Yazd human foreskin fibroblast lines (#8, #17, and #18) displaying similar characteristics to mesenchymal stromal cells: A lab resources report.
已建立的显示出与间充质基质细胞相似特征的亚兹德人包皮成纤维细胞系（#8、#17和#18）：一份实验室资源报告。
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The effect of the human cumulus cells-conditioned medium on in vitro maturation of mouse oocyte: An experimental study.人卵丘细胞条件培养基对小鼠卵母细胞体外成熟的影响：一项实验研究。
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