Abstract：
Glioma cells switch between energetic pathways to adapt and resist therapies. We present a protocol for measuring mitochondrial and glycolytic ATP rates in patient-derived glioma stem-like cells using a Seahorse XF ATP rate assay. We describe steps for growing 3D glioma stem-like cells, attaching cells to the assay plate, preparing drugs, and running the ATP rate assay. We also detail procedures for imaging viable cell numbers and normalization, with tips to overcome pitfalls in Agilent Seahorse assays.
摘要：
胶质瘤细胞在能量通路之间切换以适应和抵抗治疗。我们提出了一种使用海马XFATP率测定法测量患者衍生的神经胶质瘤干细胞样细胞中线粒体和糖酵解ATP率的方案。我们描述了生长3D神经胶质瘤干细胞样细胞的步骤，将细胞附着到测定板上，准备药物,并运行ATP速率测定。我们还详细介绍了成像活细胞数和归一化的程序，提示克服安捷伦海马检测中的陷阱。
