Single-cell assessment of the modulation of macrophage activation by ex vivo intervertebral discs using impedance cytometry.
Single-cell assessment of the modulation of macrophage activation by ex vivo intervertebral discs using impedance cytometry.
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DOI：
10.1016/j.bios.2022.114346
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发表时间：
2022-08-15
影响因子：
12.6
通讯作者：
中科院分区：
文献类型：
--
作者：
巨噬细胞活化及其对免疫调节的调节的测量对于阻止与引起慢性背痛的椎间盘变性相关的炎症反应以及与面临免疫排斥的移植相关的炎症反应具有极大的意义。由于巨噬细胞具有多种免疫功能的表型可塑性，净疾病结果由具有竞争功能的亚群的平衡决定，突出了对单细胞方法的需求，以量化其活化表型的异质性。然而，由于巨噬细胞活化可以遵循几种信号传导途径，因此用抗体对标记物进行荧光染色后的细胞计数通常不能提供关于活化动力学的剂量依赖性信息。我们提出了高通量单细胞阻抗细胞术，用于多参数测量单个巨噬细胞的生物物理变化，以剂量和持续时间依赖性的方式定量激活，而不依赖于特定的信号传导途径。在两个频率下测量的阻抗相位度量和来自较低频率下的阻抗幅值的电直径被串联使用，以通过退化盘对来自不同剂量和持续时间水平的脂多糖刺激的巨噬细胞活化进行基准，从而可以确定姜黄素的活化状态的逆转。这种无标记的单细胞测量方法可以为平台筛选炎症疗法奠定基础，从而解决背痛的慢性问题。
Measurement of macrophage activation and its modulation for immune regulation is of great interest to arrest inflammatory responses associated with degeneration of intervertebral discs that cause chronic back pain, and with transplants that face immune rejection. Due to the phenotypic plasticity of macrophages that serve multiple immune functions, the net disease outcome is determined by a balance of subpopulations with competing functions, highlighting the need for single-cell methods to quantify heterogeneity in their activation phenotypes. However, since macrophage activation can follow several signaling pathways, cytometry after fluorescent staining of markers with antibodies does not often provide dose-dependent information on activation dynamics. We present high throughput single-cell impedance cytometry for multiparametric measurement of biophysical changes to individual macrophages for quantifying activation in a dose and duration dependent manner, without relying on a particular signaling pathway. Impedance phase metrics measured at two frequencies and the electrical diameter from impedance magnitude at lower frequencies are used in tandem to benchmark macrophage activation by degenerated discs against that from lipopolysaccharide stimulation at varying dose and duration levels, so that reversal of the activation state by curcumin can be ascertained. This label-free single-cell measurement method can form the basis for platforms to screen therapies for inflammation, thereby addressing the chronic problem of back pain.
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影响因子：
9.5
作者：
Li T;Xiao L;Yang J;Ding M;Zhou Z;LaConte L;Jin L;Dorn HC;Li X
通讯作者：
Li X
影响因子：
6.4
作者：
An, HZ;Yu, YH;Cao, XT
通讯作者：
Cao, XT
影响因子：
4.8
作者：
Jacobs, AT;Ignarro, LJ
通讯作者：
Ignarro, LJ
DOI：
10.1073/pnas.1711872115
发表时间：
2018-03-20
影响因子：
11.1
作者：
Pavillon, Nicolas;Hobro, Alison J.;Smith, Nicholas I.
通讯作者：
Smith, Nicholas I.
影响因子：
6.1
作者：
Honrado C;Bisegna P;Swami NS;Caselli F
通讯作者：
Caselli F