用于秀丽隐杆线虫毒理学实验的类桥式微流控芯片平台  

An bridge-like microfluidic chip based platform for toxicity testing of Caenorhabditis elegans

作　　者：李栋基 吴佳颖 李闯[1] 高林耿 何希文 杨光[1] 栾洋[2] 陈翔[1] LI Dong-ji;WU Jia-ying;LI Chuang;GAO Lin-geng;HE Xi-wen;YANG Guang;LUAN Yang;CHEN Xiang(National Key Laboratory of Science and Technology on Micro/Nano Fabrication,Department of Micro/Nano Electronics,Shanghai Jiao Tong University,Shanghai 200240,China;Hongqiao International Institute of Medicine,Faculty of Public Health,Shanghai Jiao Tong University School of Medicine,Shanghai 200336,China)

机构地区：[1]上海交通大学微纳电子学系微米/纳米加工技术国家级重点实验室,上海200240 [2]上海交通大学医学院公共卫生学院虹桥国际医学研究院,上海200336

出　　处：《药物分析杂志》2020年第7期1160-1168,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：上海市科委基础重大研发计划资助项目(17JC1401001);上海交通大学医工交叉重点项目(ZH2018ZDA22)。

摘　　要：目的:为了解决传统微孔板中线虫毒理实验的试剂消耗量大、人工操作烦琐等问题,搭建半自动化微流控实验平台,进行秀丽隐杆线虫(Caenorhabditis elegans,简称线虫)药物毒性试验。方法:设计并使用MEMS技术制作了一种类桥式微流控芯片,并基于该芯片搭建了一个半自动化微流控平台。以一种有机磷农药久效磷(MCP)作为工具药物,考察平台对于毒性评价研究的可行性。结果:该芯片可将有限条的线虫限制于微流控芯片腔室内而不影响其正常生长运动,由于对腔室的高度做了限制,减少了线虫在Z轴方向上重叠的情况,便于后续的观察和图像处理。基于芯片的微流控平台可以半自动换液以维持药物浓度稳定,并半自动化地保存视频信息。经测试,此微流控平台可实现线虫进样和片上长期培养,结果与传统微孔板培养体系无差异;芯片内毒性测试结果显示MCP对线虫具有明显的生长发育毒性和神经毒性,与经典线虫毒性实验的结果基本一致,表明其可用于药物和线虫相互作用研究。结论:该研制平台的优势在于能实现定时半自动化换液,试剂消耗量与传统微孔板试验相比减少约50%,且便于图像分析线虫生理信息,后期可应用于药物高通量筛选和大量复杂环境化学物的复合暴露毒性测试。Objective:To solve the problems of large reagent consumption and complex manual operation in traditional worm assays in multi-well plates,an semi-automatic microfluidic experiment platform was developed for the drug toxicity test of Caenorhabditis elegans(hereinafter referred to as nematodes). Methods:A bridgelike microfluidic chip was designed and constructed using MEMS technology,and an semi-automatic microfluidic platform was built. An organophosphorus pesticide monocrotophos(MCP) was used to investigate the feasibility of the platform for drug toxicity studies based on nematodes. Results:The bridge-like chip could limit the scope of nematodes to the microfluidic chip chamber without affecting its normal growth and movement,reducing the overlap of the nematodes in the Z-axis direction with the advantage of limiting the height of the chamber,which facilitates subsequent observation and image processing. The chip-based microfluidic platform could semi-automatically change fluids to maintain drug concentration stability and capture video for further analysis. Results showed that this microfluidic experiment platform could realize worm loading and long-term culture on chip,which is the same as the traditional multi-well plates. Moreover,the on-chip drug toxicity tests showed that MCP has obvious growth and development toxicity,as well as neurotoxicity to the nematodes. The dose-response relationship was basically consistent with the results of traditional nematode toxicity experiments,indicating that this platform can be used for drug toxicity studies of nematodes. Conclusion:The advantage of this platform is that it could realize semiautomatic liquid flowing in the chamber. The reagent consumption is reduced to 50% compared with the traditional methods using multi-well plates. It can be applied to the high-throughput screening of drug toxicity and the compound exposure toxicity tests of a large number of complex environmental chemicals in the future.

关 键 词：秀丽隐杆线虫 微流控 半自动化平台 长期培养 毒理实验 高通量药物筛选 

分 类 号：R917[医药卫生—药物分析学]