集成化六通道微流控细胞迁移芯片的研制及应用

Development and application of integrated high-throughput microfluidic cell migration chip

目的 探究解决当前细胞迁移研究方法中通量低、难以建立稳定的浓度梯度并实时观察细胞迁移行为等问题。方法 该文设计了一款六通道阵列微流控芯片。首先,使用有限元分析软件COMSOL Multiphysics 5.5进行多物理场建模和数值模拟以分析细胞迁移主管道的流动行为;然后,通过测试健康人中性粒细胞在该微流控芯片中对于不同类型趋化因子梯度的趋化反应来验证该设备的通量优势;接下来,通过分析6例Ⅱ型糖尿病患者和3例健康人的中性粒细胞的迁移速度,进一步验证了环形六通道阵列微流控芯片的临床适用性;最后,使用Pearson系数分析糖尿病患者中性粒细胞趋化功能与患者部分生理指标的相关性。结果 仿真软件模拟管道内部的浓度梯度数据与管道实时荧光测试数据相拟合;健康人中性粒细胞在100 nmol/L白细胞介素-8(IL-8)环境下,其平均迁移速度为(0.21±0.01)μm/s,在100 nmol/L N-甲酰-L-甲硫氨酰-L-白氨酰-L-苯丙氨酸(fMLP)环境下为(0.22±0.01)μm/s;在健康人与糖尿病患者中性粒细胞迁移实验对比中,健康人中性粒细胞趋化速度为(0.19±0.01)μm/s,糖尿病患者中性粒细胞趋化速度为(0.15±0.02)μm/s;相关性分析表明Ⅱ型糖尿病患者中性粒细胞迁移速度与糖化血红蛋白呈负相关。结论 该文所设计的高通量微流控芯片能快速、稳定地检测细胞的迁移功能,可以作为细胞迁移研究的新工具。

Objective To solve the problems of low throughput of current cell migration research methods,which was difficult to establish a stable concentration gradient and observe cell migration behavior in real time,a six-channel array microfluidic chip was designed in this paper.Methods In this paper,a six-channel array microfluidic chip is designed.Firstly,multiphysics modeling and numerical simulation were performed using the finite element analysis software COMSOL Multiphysics 5.5 to analyze the flow behavior of the main pipeline of cell migration.Then,the throughput advantage of the device was verified by testing the chemotaxis response of healthy human neutrophils to different types of chemokine gradients in this microfluidic chip.Subsequently,by analyzing the migration rate of neutrophils in 6 patients with type II diabetes mellitus and 3 healthy people,the clinical applicability of the annular six-channel array microfluidic chip was further verified.Finally,the Pearson coefficient was used to analyze the correlation between neutrophil chemotaxis function and some physiological indicators in patients with diabetes.Results The concentration gradient data inside the pipeline simulated by the simulation software was compatible with the real-time fluorescence test data of the pipeline.The average migration rate of healthy human neutrophils was(0.21±0.01)μm/s in 100 nmol/L interleukin-8(IL-8)environment and(0.22±0.01)μm/s in 100 nmol/L N-Formyl-Met-Leu-Phe(fMLP)environment.In the comparison of neutrophil migration experiments between healthy people and diabetic patients,the chemotaxis rate of neutrophils in healthy people was(0.19±0.01)μm/s,and the neutrophil chemotaxis rate in diabetic patients was(0.15±0.02)μm/s.Correlation analysis showed that neutrophil migration rate in patients with type II diabetes mellitus was inversely correlated with glycated hemoglobin.Conclusion The high-throughput microfluidic chip proposed in this paper allowed rapid and selective detection of cell migration characteristics at the single-cell level,and it could be used as a new tool for cell migration research.