Department of Physical and Analytical Chemistry, Faculty of Chemistry, University of Oviedo, C/Julián Clavería 8, 33006 Oviedo, Spain.
Health Research Institute of the Principality of Asturias (ISPA), Avda. Hospital Universitario s/n, 33011 Oviedo, Spain.
Anal Chem. 2023 Aug 15;95(32):11874-11878. doi: 10.1021/acs.analchem.3c02506. Epub 2023 Aug 3.
The combined use of fluorescence-activated cell sorting (FACS) and single-cell inductively coupled plasma mass spectrometry (SC-ICP-MS) is reported, for the first time, in this work. It is applied to evaluate the differences between the cellular uptake of ultrasmall iron oxide nanoparticles (FeNPs) loaded with cisplatin(IV) prodrug (FeNPs-Pt(IV)) and cisplatin regarding cell viability. For this aim, FACS is applied to separate viable, apoptotic, and necrotic A2780 ovarian cancer cells after exposing them to the nanotransported prodrug and cisplatin, respectively. The different sorted cell populations are individually analyzed using quantitative SC-ICP-MS to address the intracellular amount of Pt. The highest Pt intracellular content occurs in the apoptotic cell population (about 2.1 fg Pt/cell) with a narrow intercellular distribution when using FeNPs-Pt(IV) nanoprodrug and containing the largest number of cells (75% of the total). In the case of the cisplatin-treated cells, the highest Pt content (about 1.6 fg Pt/cell) could be determined in the viable sorted cell population. The combined methodology, never explored before, permits a more accurate picture of the effect of the intracellular drug content together with the cell death mechanisms associated with the free drug and the nanotransported prodrug, respectively, and opens the door to many possible single-cell experiments in sorted cell populations.
本文首次报道了将荧光激活细胞分选（FACS）与单细胞电感耦合等离子体质谱（SC-ICP-MS）联合使用，用于评估载顺铂（IV）前药的超小氧化铁纳米颗粒（FeNPs-Pt（IV））和顺铂在细胞摄取方面的差异，以及对细胞活力的影响。为此，FACS 被用于分别分离暴露于纳米转运前药和顺铂的 A2780 卵巢癌细胞中的活细胞、凋亡细胞和坏死细胞。使用定量 SC-ICP-MS 对不同分选的细胞群进行单独分析，以确定细胞内 Pt 的含量。当使用 FeNPs-Pt（IV）纳米前药时，凋亡细胞群体中 Pt 的细胞内含量最高（约 2.1 fg Pt/细胞），细胞内分布较窄（约占总数的 75%）。在顺铂处理的细胞中，可在活细胞群体中检测到最高的 Pt 含量（约 1.6 fg Pt/细胞）。这种联合方法以前从未被探索过，它可以更准确地了解细胞内药物含量与游离药物和纳米转运前药分别相关的细胞死亡机制的影响，并为在分选细胞群体中进行许多可能的单细胞实验开辟了道路。
