基于细胞的磷脂蓄积检测成像分析方法。
Cell-Based Imaging Assay for Detection of Phospholipidosis.
机构信息
National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.
出版信息
Methods Mol Biol. 2022;2474:73-82. doi: 10.1007/978-1-0716-2213-1_8.
Accumulation of lysosomal phospholipids in cells exposed to cationic amphiphilic drugs is characteristic of drug-induced phospholipidosis. The morphological hallmark of phospholipidosis is the appearance of unicentric or multicentric-lamellar bodies when viewed under an electron microscope (EM). The EM method, the gold standard of detecting cellular phospholipidosis, has downsides, namely, low-throughput, high-costs, and unsuitability for screening a large chemical library. This chapter describes a cell-based high-content phospholipidosis assay using the LipidTOX reagent in a high-throughput screening (HTS) platform. This assay has been optimized and validated in HepG2 and HepRG cells, and miniaturized into a 1536-well plate, thus can be used for high-throughput screening (HTS) to identify chemical compounds that induce phospholipidosis.
细胞内溶酶体磷脂的积累是阳离子型两亲性药物引起磷脂蓄积症的特征。电镜（EM）下观察到的磷脂蓄积症的形态学特征是单中心或多中心层状体的出现。作为检测细胞内磷脂蓄积症的金标准，电镜法存在一些缺点，如通量低、成本高、不适合筛选大型化学文库。本章描述了一种基于细胞的高内涵磷脂蓄积症检测方法，该方法使用 LipidTOX 试剂在高通量筛选（HTS）平台上进行。该检测方法已在 HepG2 和 HepRG 细胞中进行了优化和验证，并将其小型化为 1536 孔板，因此可用于高通量筛选（HTS），以鉴定诱导磷脂蓄积症的化学化合物。
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