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接种了RTgill-W1细胞的微流控生物芯片的长期储存和基于阻抗的水毒性测试能力。

Long-term storage and impedance-based water toxicity testing capabilities of fluidic biochips seeded with RTgill-W1 cells.

作者信息

Brennan Linda M, Widder Mark W, Lee Lucy E J, van der Schalie William H

机构信息

US Army Center for Environmental Health Research, Fort Detrick, MD, USA.

出版信息

Toxicol In Vitro. 2012 Aug;26(5):736-45. doi: 10.1016/j.tiv.2012.03.010. Epub 2012 Mar 24.

DOI:10.1016/j.tiv.2012.03.010
PMID:22469871
Abstract

Rainbow trout gill epithelial cells (RTgill-W1) are used in a cell-based biosensor that can respond within one hour to toxic chemicals that have the potential to contaminate drinking water supplies. RTgill-W1 cells seeded on enclosed fluidic biochips and monitored using electric cell-substrate impedance sensing (ECIS) technology responded to 18 out of the 18 toxic chemicals tested within one hour of exposure. Nine of these chemical responses were within established concentration ranges specified by the U.S. Army for comparison of toxicity sensors for field application. The RTgill-W1 cells remain viable on the biochips at ambient carbon dioxide levels at 6°C for 78weeks without media changes. RTgill-W1 biochips stored in this manner were challenged with 9.4μM sodium pentachlorophenate (PCP), a benchmark toxicant, and impedance responses were significant (p<0.001) for all storage times tested. This poikilothermic cell line has toxicant sensitivity comparable to a mammalian cell line (bovine lung microvessel endothelial cells (BLMVECs)) that was tested on fluidic biochips with the same chemicals. In order to remain viable, the BLMVEC biochips required media replenishments 3 times per week while being maintained at 37°C. The ability of RTgill-W1 biochips to maintain monolayer integrity without media replenishments for 78weeks, combined with their chemical sensitivity and rapid response time, make them excellent candidates for use in low cost, maintenance-free field-portable biosensors.

摘要

虹鳟鱼鳃上皮细胞(RTgill-W1)被用于一种基于细胞的生物传感器,该传感器能够在一小时内对有可能污染饮用水供应的有毒化学物质做出反应。接种在封闭流体生物芯片上并使用细胞-基质阻抗传感(ECIS)技术进行监测的RTgill-W1细胞,在接触测试的18种有毒化学物质中的18种后,一小时内就做出了反应。其中九种化学物质的反应在美军规定的用于野外应用毒性传感器比较的既定浓度范围内。RTgill-W1细胞在生物芯片上于6°C的环境二氧化碳水平下可存活78周,无需更换培养基。以这种方式储存的RTgill-W1生物芯片用9.4μM五氯酚钠(PCP)(一种基准毒物)进行挑战,在所有测试的储存时间内,阻抗反应均显著(p<0.001)。这种变温细胞系对毒物的敏感性与在相同化学物质的流体生物芯片上测试的哺乳动物细胞系(牛肺微血管内皮细胞(BLMVECs))相当。为了保持存活,BLMVEC生物芯片在维持于37°C时每周需要补充三次培养基。RTgill-W1生物芯片能够在不补充培养基的情况下保持单层完整性达78周,再加上其化学敏感性和快速反应时间,使其成为用于低成本、无需维护的现场便携式生物传感器的极佳候选者。

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