基于微流控技术的细胞水平高通量药物筛选系统的研究进展

doi:10.3724/SP.J.1123.2020.07014

摘要/Abstract

摘要：

药物筛选是新药研发的关键步骤,创新药物的发现需要采用适当的药物作用靶点对大量化合物样品进行筛选。高通量筛选系统能够实现数千个反应同时测试和分析,大大提高了药物筛选的实验规模和效率。其中基于细胞水平的高通量药物筛选系统因为更加接近人体生理条件,成为主要的筛选模式。而目前发展成熟的高通量细胞筛选系统主要基于多孔板,存在细胞培养条件单一、耗时费力、试剂消耗量大等问题,且较难实现复杂的组合药物筛选。微流控技术作为一种在微米尺度通道中操纵和控制微流体的技术,具有微量、高效、高通量和自动化的优点,能较好地克服多孔板筛选系统的不足,为构建细胞高通量药物筛选系统提供了一种高效、可靠的技术手段。微流控系统在细胞培养材料、芯片结构设计和流体控制方面均可灵活变化,能更好地实现对细胞生长微环境的调控和模拟。文章综述了基于微流控技术的细胞水平高通量药物筛选系统的研究进展,按照不同的微流体操控模式,对基于灌注流、液滴和微阵列的3种类型的微流控细胞筛选系统进行了分类介绍,并分别总结了它们的优缺点,最后展望了微流控细胞水平高通量药物筛选系统的发展前景,提出了该领域目前存在的问题以及解决问题的方向。

关键词: 微流控技术, 药物筛选, 高通量筛选, 综述

Abstract:

Drug screening is the process of screening new drugs or leading compounds with biological activity from natural products or synthetic compounds, and it plays an essential role in drug discovery. The discovery of innovative drugs requires the screening of a large number of compounds with appropriate drug targets. With the development of genomics, proteomics, metabolomics, combinatorial chemistry, and other disciplines, the library of drug molecules has been largely expanded, and the number of drug targets is continuously increasing. High-throughput screening systems enable the parallel analysis of thousands of reactions through automated operation, thereby enhancing the experimental scale and efficiency of drug screening. Among them, cell-based high-throughput drug screening has become the main screening mode because it can provide a microenvironment similar to human physiological conditions. However, the current high-throughput screening systems are mainly built based on multiwell plates, which have several disadvantages such as simple cell culture conditions, laborious and time-consuming operation, and high reagent consumption. In addition, it is difficult to achieve complex drug combination screening. Therefore, there is an urgent need for rapid and low-cost drug screening methods to reduce the time and cost of drug development. Microfluidic techniques, which can manipulate and control microfluids in microscale channels, have the advantages of low consumption, high efficiency, high throughput, and automation. It can overcome the shortcomings of screening systems based on multi-well plates and provide an efficient and reliable technical solution for establishing high-throughput cell-based screening systems. Moreover, microfluidic systems can be flexibly changed in terms of cell culture materials, chip structure design, and fluid control methods to enable better control and simulation of cell growth microenvironment. Operations such as cell seeding, culture medium replacement or addition, drug addition and cleaning, and cell staining reagent addition are usually involved in cell-based microfluidic screening systems. These operations are all based on the manipulation of microfluids. This paper reviews the research advances in cell-based microfluidic screening systems using different microfluidic manipulation modes, namely perfusion flow mode, droplet mode, and microarray mode. In addition, the advantages and disadvantages of these systems are summarized. Moreover, the development prospects of high-throughput screening systems based on microfluidic techniques has been looked forward. Furthermore, the current problems in this field and the directions to overcome these problems are discussed.

Key words: microfluidic technique, drug screening, high throughput screening, review

中图分类号:

O658

梁怡萧, 潘建章, 方群. 基于微流控技术的细胞水平高通量药物筛选系统的研究进展[J]. 色谱, 2021, 39(6): 567-577.

LIANG Yixiao, PAN Jianzhang, FANG Qun. Research advances of high-throughput cell-based drug screening systems based on microfluidic technique[J]. Chinese Journal of Chromatography, 2021, 39(6): 567-577.

图/表 6

图 1 基于灌注流模式的微流控细胞筛选系统

Fig. 1 Cell-based microfluidic screening systems based on perfusion flow mode a. schematic diagram of the pneumatically-actuated elastomeric valves based microfluidic platform for cell cytotoxicity screening[17]; b. schematic diagram of the microfluidic system based on bridge-and-underpass architecture[27]; c. schematic diagram of the integrated microfluidic device with concentration gradient generators[28]; d. schematic diagram of the 3D printed concentration gradient generator[32].

图 2 基于3D细胞培养的灌注流模式微流控细胞筛选系统

Fig. 2 3D cell culture-based microfluidic screening systems based on the perfusion flow mode a. schematic diagram of cell chambers with two sizes used for the formation of cell spheres[38]; b. schematic diagram of the 3D cell perfusion culture chip with an array of micropillars in the microfluidic channel[39]; c. schematic diagram of the integrated microchip with interdigitated microelectrodes[20].

图 3 基于灌注流模式的器官芯片系统

Fig. 3 Organ-on-a-chip systems based on perfusion flow mode a. schematic diagram of the lung-on-a-chip system[42]; b. schematic diagram of the organ-on-a-chip system for studying tumor-induced angiogenesis[49]; c. schematic diagram of the organ-on-a-chip system integrating four types of organ cells[50].

图 4 基于液滴模式的微流控细胞筛选系统

Fig. 4 Cell-based microfluidic screening systems based on droplet mode a. workflow of the droplet-based drug screening platform[54]; b. schematic diagram of the microwell array for randomly pairing droplets[55]; c. schematic diagram of the microchannel chamber for trapping droplets[56].

图 5 基于3D细胞培养的液滴模式微流控细胞筛选系统

Fig. 5 Droplet mode microfluidic systems based on 3D cell culture a. schematic diagram of the microfluidic flow-focusing device for generating mixed hydrogel droplets[57]; b. formation of coculture tumor spheroid with stromal fibroblast cells[58]; c. illustration of cell culture and drug testing process in the sidewall-attached droplet array: Fig. c1. cell seeding; Fig. c2. formation of cell spheroid in a droplet; Fig. c3. addition of the drug; Fig. c4. addition of fluorescence dye to stain the cell spheroid[62].

图 6 基于微阵列模式的微流控细胞筛选系统

Fig. 6 Cell-based microfluidic screening systems based on microarray mode a. schematic diagram of the DataChip platform for testing of compound toxicity[63]; b. schematic diagram of the cell-seeded microwell and chemical-laden post aligned and sandwiched together[68]; c. schematic diagram of the liver-immune coculture array chip[71]; d. formation of the cell droplet array based on superhydrophilic-superhydrophobic micropattern[75]; e. illustration of the drug combination screening based on sequential operation droplet array (SODA) system[77].

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