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用于精确测量全血滴中小鼠、大鼠和人类中性粒细胞趋化性的微流控分析方法。

Microfluidic assay for precise measurements of mouse, rat, and human neutrophil chemotaxis in whole-blood droplets.

作者信息

Jones Caroline N, Hoang Anh N, Martel Joseph M, Dimisko Laurie, Mikkola Amy, Inoue Yoshitaka, Kuriyama Naohide, Yamada Marina, Hamza Bashar, Kaneki Masao, Warren H Shaw, Brown Diane E, Irimia Daniel

机构信息

BioMEMS Resource Center, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA;

Center for Comparative Medicine, Massachusetts General Hospital, Boston, Massachusetts, USA.

出版信息

J Leukoc Biol. 2016 Jul;100(1):241-7. doi: 10.1189/jlb.5TA0715-310RR. Epub 2016 Jan 27.

Abstract

Animal models of human disease differ in innate immune responses to stress, pathogens, or injury. Precise neutrophil phenotype measurements could facilitate interspecies comparisons. However, such phenotype comparisons could not be performed accurately with the use of current assays, as they require the separation of neutrophils from blood using species-specific protocols, and they introduce distinct artifacts. Here, we report a microfluidic technology that enables robust characterization of neutrophil migratory phenotypes in a manner independent of the donor species and performed directly in a droplet of whole blood. The assay relies on the particular ability of neutrophils to deform actively during chemotaxis through microscale channels that block the advance of other blood cells. Neutrophil migration is measured directly in blood, in the presence of other blood cells and serum factors. Our measurements reveal important differences among migration counts, velocity, and directionality among neutrophils from 2 common mouse strains, rats, and humans.

摘要

人类疾病的动物模型在对应激、病原体或损伤的先天免疫反应方面存在差异。精确的中性粒细胞表型测量有助于进行种间比较。然而,使用当前的检测方法无法准确进行此类表型比较,因为它们需要使用物种特异性方案从血液中分离中性粒细胞,并且会引入明显的假象。在此,我们报告一种微流控技术,该技术能够以独立于供体物种的方式并直接在全血滴中对中性粒细胞迁移表型进行可靠表征。该检测方法依赖于中性粒细胞在趋化作用期间通过微尺度通道主动变形的特殊能力,这些微尺度通道会阻止其他血细胞前进。在存在其他血细胞和血清因子的情况下,直接在血液中测量中性粒细胞迁移。我们的测量揭示了来自两种常见小鼠品系、大鼠和人类的中性粒细胞在迁移计数、速度和方向性方面的重要差异。

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