主要人类颗粒细胞中自由基的量子感知与纳米尺度分辨率
Quantum Sensing of Free Radicals in Primary Human Granulosa Cells with Nanoscale Resolution
作者:Nuan Lin;Koen van Zomeren;Teelkien van Veen;Aldona Mzyk;Yue Zhang;Xiaoling Zhou;Torsten Plosch;Uwe J. F. Tietge;Astrid Cantineau;Annemieke Hoek;Romana Schirhagl;
DOI:https://doi.org/10.1021/acscentsci.3c00747
发表时间:2023年
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摘要
颗粒层细胞(cGCs)和壁层颗粒细胞(mGCs)虽然来源于相同的前体细胞,但在解剖和功能上是异质的。它们通过支持卵母细胞的能力和卵泡发育对女性生育力至关重要。有各种技术可用于研究mGCs和cCGs中自由基的作用。然而,时间和空间分辨率仍然是一个挑战。我们采用量子传感方法研究了从进行体外受精(IVF)的女性中分离出的cGCs和mGCs中的纳米尺度自由基的产生。细胞被孵育在裸的荧光纳米金刚石(FNDs)或线粒体定位的FNDs中,以检测细胞质和线粒体中的自由基。在用menadione诱导氧化应激后,我们继续检测了30分钟的自由基产生。我们观察到cGCs和mGCs中的自由基产生在10分钟后增加。虽然在cGCs和mGCs的生理状态下,细胞质和线粒体中的自由基水平无法区分,但我们测得的线粒体中的自由基变化在两种细胞类型中都显著更大,表明线粒体是自由基产生的地点。此外,我们观察到cGCs中的细胞质自由基变化发生较晚,且比例较小,表明cGCs可能对氧化应激更具抵抗力。
Abstract
Cumulus granulosa cells (cGCs) and mural granulosa cells (mGCs), although derived from the same precursors, are anatomically and functionally heterogeneous. They are critical for female fertility by supporting oocyte competence and follicular development. There are various techniques used to investigate the role of free radicals in mGCs and cCGs. Yet, temporospatial resolution remains a challenge. We used a quantum sensing approach to study free radical generation at nanoscale in cGCs and mGCs isolated from women undergoing oocyte retrieval during in vitro fertilization (IVF). Cells were incubated with bare fluorescent nanodiamonds (FNDs) or mitochondria targeted FNDs to detect free radicals in the cytoplasm and mitochondria. After inducing oxidative stress with menadione, we continued to detect free radical generation for 30 min. We observed an increase in free radical generation in cGCs and mGCs from 10 min on. Although cytoplasmic and mitochondrial free radical levels are indistinguishable in the physiological state in both cGCs and mGCs, the free radical changes measured in mitochondria were significantly larger in both cell types, suggesting mitochondria are sites of free radical generation. Furthermore, we observed later occurrence and a smaller percentage of cytoplasmic free radical change in cGCs, indicating that cGCs may be more resistant to oxidative stress.
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