睾丸细胞衍生条件培养基支持人类胚胎干细胞的生殖细胞分化

IF 16.4 1区 化学 Q1 CHEMISTRY, MULTIDISCIPLINARY
Fatemeh Akyash, Reza Aflatoonian, Ehsan Farashahi-Yazd, Fatemeh Hajizadeh-Tafti, Jalal Golzadeh, Somayyeh Sadat Tahajjodi, Behrouz Aflatoonian
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引用次数: 0

摘要

目的:人类生殖细胞发育研究面临伦理和技术挑战。因此,本研究旨在体外分化作为多能细胞的人类胚胎干细胞(hESCs)为生殖细胞,这是研究配子发生的分子和细胞方面以及了解不育原因的宝贵工具:在这项实验研究中,两种不同的完全培养基[杜氏改良老鹰培养基(Dulbecco's Modified Eagle Medium,DMEM)+20%胎牛血清(FBS)和胚状体(EBs)培养基;不含碱性成纤维细胞生长因子(bFGF)的KOSR/HES]分别用于使用睾丸细胞条件培养基(TCCM)的试验组和自发分化(SD)的对照组。因此,将来自 hESCs(Yazd2;46XY)的 EB 在不同条件下培养:EB 培养基;EB 培养基和条件 EB 培养基;未经条件培养的 EB 培养基、DMEM 和 FBS;EB 培养基、条件 DMEM 和 FBS 培养基。4天、7天和14天后收集EB,使用IF和相对反转录定量聚合酶链反应(RT-qPCR)评估其基因表达谱,并与第0天的hESCs进行比较:结果:与 SD 组相比,TCCM 组的配子发生基因表达水平有所提高。此外,各组分化细胞的免疫染色均显示体外配子生成(IVG):我们的研究结果表明,人类 TCCM 可作为男性和女性生殖细胞体外发育的天然龛位。结论:我们的研究结果表明,人类 TCCM 可作为男性和女性生殖细胞体外发育的天然龛位,但还需要进一步研究,以确定人类 TCCM 中的各种因素和代谢物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Testicular Cells Derived Conditioned Medium Supports Germ Cell Differentiation of Human Embryonic Stem Cells.

Objective: There are ethical and technical challenges in studying human germ cell development. Therefore, the aim of the study is in vitro differentiation of human embryonic stem cells (hESCs), as pluripotent cells, to the germ cells which is a valuable tool for studying molecular and cellular aspects of gametogenesis and understanding causes of infertility.

Materials and methods: In this experimental study, two different complete media [Dulbecco's Modified Eagle Medium (DMEM)+20% fetal bovine serum (FBS) and embryoid bodies (EBs) medium; KOSR/HES without basic fibroblast growth factor (bFGF)] were used in the both of test groups using testicular cells derived conditioned medium (TCCM) and control groups spontaneously differentiated (SD). Thereby, EBs from hESCs (Yazd2; 46XY) were cultured in different conditions EB medium; EB medium and conditioned EB medium; EB medium, DMEM, and FBS without conditioning; EB medium, conditioned DMEM, and FBS medium. EBs were collected after 4, 7, and 14 days and their gene expression profiles were assessed and compared to hESCs, as day 0, using IF and relative reverse transcription quantitative polymerase chain reaction (RT-qPCR).

Results: An increase in the gametogenesis gene expression level in TCCM groups was showed in comparison with SD groups. Additionally, immunostaining of differentiated cells in all groups showed in vitro gametogenesis (IVG).

Conclusion: Our findings showed that human TCCM could be used as a natural niche for in vitro male and female germ cell development. However, further studies are needed to define the factors and metabolites within the human TCCM.

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来源期刊
Accounts of Chemical Research
Accounts of Chemical Research 化学-化学综合
CiteScore
31.40
自引率
1.10%
发文量
312
审稿时长
2 months
期刊介绍: Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.
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