对 THP-1 单核细胞在氧化铟锡电极上的巨噬细胞极化进行无标记和实时电阻抗监测

IF 5.5 3区 工程技术 Q1 BIOCHEMICAL RESEARCH METHODS BioChip Journal Pub Date : 2023-12-27 DOI:10.1007/s13206-023-00132-0
Duc-Trung Pham, Hoang Lan Pham, Hien T. Ngoc Le, My-Van Tieu, Daesoo Kim, Jae Young Kim, Sungbo Cho
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摘要

巨噬细胞是一种免疫细胞,在人体对病原体和肿瘤细胞的初始免疫反应中发挥着重要作用。我们研究了利用电阻抗监测来评估 THP-1 单核细胞向巨噬细胞分化的情况,这对于开展免疫疗法研究十分必要。所测得的 1 kHz 时的电阻变化和 100 kHz 时的电容变化不仅与经磷脂-12-肉豆蔻酸-13-醋酸酯处理后分化的静息巨噬细胞密度的增加成正比,而且还与播种在电极上的 THP-1 细胞的初始数量成正比。此外,THP-1 细胞培养 48 小时后的实时阻抗数据表明,与单核细胞表型(悬浮细胞)相比,静止巨噬细胞表型(粘附细胞)对微电极表面的识别率更高,阻抗信号显著增加。此外,在巨噬细胞极化阶段,交替活化巨噬细胞表型比经典活化巨噬细胞和静止巨噬细胞表型更大、更扁平,这表明在 1 kHz 和 100 kHz 频率下,交替活化巨噬细胞(4750 Ω 和 - 3.5 nF)比经典活化巨噬细胞(2000 Ω 和 - 1.5 nF)和静止巨噬细胞(3500 Ω 和 - 2.0 nF)分别具有更高的电阻和更低的电容阻抗。研究结果表明,阻抗测量系统在监测巨噬细胞分化和极化方面具有很高的灵敏度和可信度。电阻抗对每种巨噬细胞表型都有意义,它与使用流式细胞仪和显微镜观察到的巨噬细胞特征一致。
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Label-Free and Real-Time Electrical Impedance Monitoring of Macrophage Polarization of THP-1 Monocytes on Indium Tin Oxide Electrode

Macrophages are immune cells that play important roles in the human body’s initial immune responses against pathogens and tumor cells. We investigated the use of electrical impedance monitoring to assess the differentiation of THP-1 monocyte into macrophages, which is necessary for immunotherapy research conducted. The change in resistance at 1 kHz and capacitance at 100 kHz measured were proportionally increased according to not only the increase in the density of resting macrophages differentiated by Phorbol-12-myristate-13-acetate treatment but also the initial number of THP-1 cells seeded on the electrode. Additionally, real-time impedance data from THP-1 cells after 48 h of cultivation demonstrated greater recognition of the resting macrophage phenotypes (adhesion cells) covered microelectrode surface with a significant increase of impedance signal in comparison with monocytes phenotypes (suspended cells). Furthermore, during the polarization phase of macrophages, the alternatively activated macrophage phenotype was larger and flatter than that of classically activated macrophage and resting macrophage phenotypes, indicating a correlation with a higher resistance and lower capacitance impedances at 1 kHz and 100 kHz of alternatively activated macrophages (4750 Ω and – 3.5 nF) than that of classically activated macrophages (2000 Ω and – 1.5 nF) and resting macrophages (3500 Ω and – 2.0 nF), respectively. The study’s findings demonstrated that the impedance measurement system is high sensitivity and confidence in monitoring macrophages differentiation and polarization. The electrical impedance, which has significance for each macrophage phenotype, is compatible with macrophages characteristic features observed using flow cytometry and a microscope.

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来源期刊
BioChip Journal
BioChip Journal 生物-生化研究方法
CiteScore
7.70
自引率
16.30%
发文量
47
审稿时长
6-12 weeks
期刊介绍: BioChip Journal publishes original research and reviews in all areas of the biochip technology in the following disciplines, including protein chip, DNA chip, cell chip, lab-on-a-chip, bio-MEMS, biosensor, micro/nano mechanics, microfluidics, high-throughput screening technology, medical science, genomics, proteomics, bioinformatics, medical diagnostics, environmental monitoring and micro/nanotechnology. The Journal is committed to rapid peer review to ensure the publication of highest quality original research and timely news and review articles.
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