使用 Seahorse XFe24 细胞外通量分析仪实时评估滋养细胞和合体滋养层细胞中的线粒体功能。

Real-Time Assessment of Mitochondrial Function in Cytotrophoblast and Syncytialized Trophoblast Cells Using the Seahorse XFe24 Extracellular Flux Analyzer.

机构信息

Graduate Programme in Medical Sciences, McMaster University, Hamilton, ON, Canada.

Department of Pediatrics, McMaster University, Hamilton, ON, Canada.

出版信息

Methods Mol Biol. 2024;2728:137-147. doi: 10.1007/978-1-0716-3495-0_12.

DOI:10.1007/978-1-0716-3495-0_12

PMID:38019398

Abstract

Physiological processes utilize variable amounts of energy required for optimal growth, development, and survival. This energy is supplied by total intracellular adenosine triphosphate (ATP) and is mainly generated by mitochondrial oxidative phosphorylation and to a lesser extent via glycolysis. Here, we provide a detailed protocol for obtaining measurements of energy metabolism using the Seahorse XFe24 Extracellular Flux Analyzer. Specifically, this assay measures mitochondrial oxidative phosphorylation based on oxygen consumption rate (OCR) and glycolysis by analyzing the extracellular acidification rate (ECAR) via real-time live cell analysis. Using trophoblast cell lines, this protocol focuses on analyzing mitochondrial respiration for both cytotrophoblasts and syncytiotrophoblasts.

摘要

生理过程利用不同数量的能量来实现最佳的生长、发育和生存。这些能量由细胞内三磷酸腺苷（ATP）总量提供，主要通过线粒体氧化磷酸化产生，其次是通过糖酵解产生。在这里，我们提供了一个详细的方案，使用 Seahorse XFe24 细胞外通量分析仪来获得能量代谢的测量结果。具体来说，该测定通过实时活细胞分析，基于耗氧量（OCR）和细胞外酸化率（ECAR）分析来测量线粒体氧化磷酸化和糖酵解。使用滋养层细胞系，该方案侧重于分析滋养细胞和合体滋养层细胞的线粒体呼吸。