摘要
细胞迁移是指细胞朝着特定的化学浓度梯度发生定向迁移运动,其在胚胎发育、伤口愈合、肿瘤转移中发挥着至关重要的作用。当前研究手段大多通量低,难以综合考虑不同浓度梯度条件对细胞迁移行为的影响。针对上述问题,本文首先设计了一款四通道微流控芯片,其特征如下:借助层流和扩散机制在细胞迁移主通道中建立和维持浓度梯度;可在单一显微镜视野下同时观测四组细胞迁移现象;集成了宽度为20μm的细胞隔离带,可校准细胞初始位置,保证实验结果的准确性。随后,借助Comsol Multiphysics有限元分析软件完成了微流控芯片的仿真分析,证明了芯片上设计S型微通道和水平压力平衡通道有助于在细胞迁移主通道中形成稳定的浓度梯度。最后,采用不同浓度(0、0.2、0.5、1.0μmol·L^(-1))与糖尿病及其并发症密切相关的晚期糖基化终末产物(AGEs)孵育中性粒细胞,研究了其在100 nmol·L^(-1)趋化因子fMLP浓度梯度环境中的迁移行为。结果表明,AGEs抑制了中性粒细胞的迁移能力,证明了四通道微流控芯片的可靠性和实用性。
Cell migration is defined as the directional movement of cells toward a specific chemical concentration gradient,which plays a crucial role in embryo development,wound healing and tumor metastasis.However,current research methods showed low flux and are only suitable for single-factor assessment,and it was difficult to comprehensively consider the effects of other parameters such as different concentration gradients on cell migration behavior.In this paper,a four-channel microfluidic chip was designed.Its characteristics were as follows:it relied on laminar flow and diffusion mechanisms to establish and maintain a concentration gradient;it was suitable for observation of cell migration in different concentration gradient environment under a single microscope field;four cell isolation zones(20μm width)were integrated into the microfluidic device to calibrate the initial cell position,which ensured the accuracy of the experimental results.In particular,we used COMSOL Multiphysics software to simulate the structure of the chip,which demonstrated the necessity of designing S-shaped microchannel and horizontal pressure balance channel to maintain concentration gradient.Finally,neutrophils were incubated with advanced glycation end products(AGEs,0,0.2,0.51.0μmol·L^(-1)),which were closely related to diabetes mellitus and its complications.The migration behavior of incubated neutrophils was studied in the 100 nmol·L^(-1) of chemokine(N-formylmethionyl-leucyl-phenyl-alanine)concentration gradient.The results prove the reliability and practicability of the microfluidic chip.
作者
李慧来
杨逍
吴晓松
李志刚
洪承刚
刘勇
朱灵
杨柯
LI Huilai;YANG Xiao;WU Xiaosong;LI Zhigang;HONG Chenggang;LIU Yong;ZHU Ling;YANG Ke(Anhui Institute of Optics and Fine Mechanics,Hefei Institutes of Physical Science,Chinese Academy of Sciences,Hefei 230031,P.R.China;University of Science and Technology of China,Hefei 230026,P.R.China;School of Biomedical Engineering,Anhui Medical University,Hefei 230032,P.R.China;Hefei Zhongke Yikangda Biomedical Co.,Ltd.,Hefei 230031,P.R.China)
出处
《生物医学工程学杂志》
EI
CAS
CSCD
北大核心
2022年第1期128-138,共11页
Journal of Biomedical Engineering
基金
国家自然科学基金(31800713)
安徽省重点研发计划(201904c03020001)
中国科学院青年创新促进会(2021442)。
关键词
微流控芯片
中性粒细胞
趋化性
晚期糖基化终末产物
抑制
Microfluidic concentration gradient chip
Neutrophils
Chemotaxis
Advanced glycation end products
Inhibition