Gao Chaoru, Yang Xiao, Liu Lijuan, Wang Yue, Zhu Ling, Zhou Jinhua, Liu Yong, Yang Ke
School of Biomedical Engineering, Anhui Medical University, Hefei 230032, P. R. China.
Anhui Institute of Optics and Precision Mechanics, Hefei Institute of Physical Sciences, Chinese Academy of Sciences, Hefei 230031, P. R. China.
Sheng Wu Yi Xue Gong Cheng Xue Za Zhi. 2023 Dec 25;40(6):1217-1226. doi: 10.7507/1001-5515.202304002.
Reduced chemotactic migration of polymorphonuclear neutrophil (PMN) in sepsis patients leads to decreased bacterial clearance and accelerates the progression of sepsis disease. Quantification of PMN chemotaxis in sepsis patients can help characterize the immune health of sepsis patients. Microfluidic microarrays have been widely used for cell chemotaxis analysis because of the advantages of low reagent consumption, near-physiological environment, and visualization of the migration process. Currently, the study of PMN chemotaxis using microfluidic chips is mainly limited by the cumbersome cell separation operation and low throughput of microfluidic chips. In this paper, we first designed an inertial cell sorting chip to achieve label-free separation of the two major cell types by using the basic principle that leukocytes (mainly granulocytes, lymphocytes and monocytes) and erythrocytes move to different positions of the spiral microchannel when they move in the spiral microchannel under different strength of inertial force and Dean's resistance. Subsequently, in this paper, we designed a multi-channel cell migration chip and constructed a microfluidic PMN inertial label-free sorting and chemotaxis analysis platform. The inertial cell sorting chip separates leukocyte populations and then injects them into the multi-channel cell migration chip, which can complete the chemotaxis test of PMN to chemotactic peptide (fMLP) within 15 min. The remaining cells, such as monocytes with slow motility and lymphocytes that require pre-activation with proliferative culture, do not undergo significant chemotactic migration. The test results of sepsis patients ( =6) and healthy volunteers ( =3) recruited in this study showed that the chemotaxis index (CI) and migration velocity ( ) of PMN from sepsis patients were significantly weaker than those from healthy volunteers. In conclusion, the microfluidic PMN inertial label-free sorting and chemotaxis analysis platform constructed in this paper can be used as a new tool for cell label-free sorting and migration studies.
脓毒症患者中多形核中性粒细胞(PMN)趋化迁移减少导致细菌清除率降低,并加速脓毒症疾病的进展。对脓毒症患者的PMN趋化性进行定量分析有助于了解脓毒症患者的免疫健康状况。微流控微阵列由于具有试剂消耗低、接近生理环境以及可观察迁移过程等优点,已被广泛用于细胞趋化性分析。目前,使用微流控芯片研究PMN趋化性主要受到细胞分离操作繁琐和微流控芯片通量低的限制。在本文中,我们首先设计了一种惯性细胞分选芯片,利用白细胞(主要是粒细胞、淋巴细胞和单核细胞)和红细胞在不同强度的惯性力和迪恩阻力作用下在螺旋微通道中移动时会移动到螺旋微通道不同位置的基本原理,实现两种主要细胞类型的无标记分离。随后,我们设计了一种多通道细胞迁移芯片,并构建了一个微流控PMN惯性无标记分选和趋化性分析平台。惯性细胞分选芯片分离白细胞群体,然后将其注入多通道细胞迁移芯片,该芯片可在15分钟内完成PMN对趋化肽(fMLP)的趋化性测试。其余细胞,如运动缓慢的单核细胞和需要通过增殖培养进行预激活的淋巴细胞,不会发生明显的趋化迁移。本研究招募的脓毒症患者(n = 6)和健康志愿者(n = 3)的测试结果表明,脓毒症患者PMN的趋化指数(CI)和迁移速度(v)明显弱于健康志愿者。总之,本文构建的微流控PMN惯性无标记分选和趋化性分析平台可作为细胞无标记分选和迁移研究的新工具。