Korangath Preethi, Ivkov Robert
Dept of Radiation Oncology and Molecular Radiation Sciences, School of Medicine, Johns Hopkins University, Baltimore, MD 21231 USA.
Department of Oncology, School of Medicine, Johns Hopkins University, Baltimore, MD 21231 USA.
Bio Protoc. 2020 Nov 20;10(22):e3822. doi: 10.21769/BioProtoc.3822.
A clear understanding of nanoparticle interactions with living systems at the cellular level is necessary for developing nanoparticle-based therapeutics. Magnetic iron oxide nanoparticles provide unique opportunities to study these interactions because of their responsiveness to magnetic fields. This enables sorting of cells containing nanoparticles from models. Once sorted, flow cytometry can identify individual cell types, which can be further analyzed for iron content, gene or protein expression changes associated with nanoparticle uptake, and for other biological responses at a molecular level. Here we provide a detailed protocol to sort and identify cells in the tumor microenvironment that have internalized magnetic iron oxide nanoparticles following intravenous administration.
为了开发基于纳米颗粒的治疗方法，有必要在细胞水平上清楚地了解纳米颗粒与生物系统的相互作用。磁性氧化铁纳米颗粒因其对磁场的响应性而提供了研究这些相互作用的独特机会。这使得能够从模型中筛选出含有纳米颗粒的细胞。筛选后，流式细胞术可以识别单个细胞类型，进而可以进一步分析其铁含量、与纳米颗粒摄取相关的基因或蛋白质表达变化，以及分子水平上的其他生物学反应。在此，我们提供了一份详细的方案，用于对静脉注射后内化了磁性氧化铁纳米颗粒的肿瘤微环境中的细胞进行分选和鉴定。
