在 CYRIS® 分析平台上使用 HEPG2 细胞进行体外检测以评估铁死亡和细胞凋亡
IN-VITRO ASSAY WITH HEPG2 CELLS ON THE CYRIS® ANALYSIS
PLATFORM TO EVALUATE FERROPTOSIS AND APOPTOSIS
作者：Christina Heichler & Dr. Peter Wolf（2022）
Abstract：
The following protocol describes the preparation, run and results of an in-vitro assay on the CYRIS® analysis platform to study ferroptotic or apoptotic processes based on the oxygen consumption rate (OCR) and morphological changes of a human hepatocarcinomaderived cell line (HepG2). The automated CYRIS® platform measures OCR and acquires microscopic images of the cells label-free in parallel and in real time. With these parameters and optional downstream analyses such as fluorescence staining and microscopy, it is possible to gain deeper insight into cell death processes. HepG2 cells are measured for 12 hours without treatment for baseline, 24 hours under treatment with common ferroptosis or apoptosis inducers and/or inhibitors and additional 6 hours after treatment washout. The results show that ferroptosis and apoptosis can be induced by different inducers such as Erastin or RSL3 (ferroptosis) and H2O2 (apoptosis). Furthermore, the ferroptosis inhibitor Ferrostatin blocks Erastin- and RSL3-induced ferroptosis. Microscopic images reveal different morphological changes of HepG2 cells after treatment with Erastin/RSL3 or H2O2. Correlations between the different cell parameters are clearly visible. It is possible and may be necessary to adapt the protocol to other cell lines, cell numbers, and culture times. It is recommended to read the entire protocol once complete before the first test procedure to avoid
文章摘要：
以下方案描述了在Cyris®分析平台上进行的体外检测的准备、运行和结果，该检测基于人肝癌细胞系(HepG2)的耗氧率(OCR)和形态变化来研究铁死亡或凋亡过程。自动化的Cyris®平台可以实时测量OCR并获取无标记细胞的显微图像。通过这些参数和可选的下游分析，如荧光染色和显微镜，可以更深入地了解细胞死亡过程。
在未处理的情况下，测量HepG2细胞12小时，在使用常见的铁死亡或细胞凋亡诱导剂和/或抑制剂处理后测量24小时，并在治疗清除后额外测量6小时。
结果表明，Erastin或RSL3(铁死亡)、H_2O_2(细胞凋亡)等不同的诱导剂均可诱导细胞发生铁死亡和细胞凋亡。另外，铁死亡抑制剂能阻断Erastin和RSL3诱导的铁死亡。显微图像揭示了用 Erastin/RSL3 或 H2O2 处理后 HepG2 细胞的不同形态变化。不同细胞参数之间的相关性清晰可见。
有可能的，也是有必要的，适应其他细胞系、细胞数量和培养时间。建议在完成第一个测试程序之前阅读整个案例，以避免
点击链接即可查看和下载文章：https://www.cyris.bio/wp-content/uploads/2022/12/20220712-Ferroptosis-Assay_Application-Note_my.pdf
文章题目、关键词与摘要译文仅用于参考。
