Microfluidic System for Automated Cell Toxicity Screening

描述（由申请人提供）：本提案的主要目的是开发用于在培养的人类细胞中进行癌症药物毒性筛选的微流体平台。虽然人们认为，关于患者个体癌症特征的改进信息可以帮助诊断和治疗，但目前可用于验证这一主张的技术是有限的。这项工作的长期目标是将微流体筛选平台商业化，以提供可用于临床环境的紧凑、低成本、自动化筛选系统。该提案的具体目标是使先前开发的微流体细胞培养阵列自动化，并证明以微流体形式进行癌症药物毒性筛选的可行性和再现性。可寻址8x3单元微流体阵列的设计和制造将利用公司内部开发的与软光刻技术相关的专业知识。通过阵列的流体输送的自动化将通过实施用工业气动接口控制的新型微流体阀来实现。将在暴露于抗癌药物（如依托泊苷）7天的HeLa细胞上收集癌细胞细胞毒性的初步证明。将在阵列中收集细胞活力以及凋亡动力学（通过荧光测定定量），并确定实验稳健性。将响应和统计均匀度与在96孔板中进行的相同试验进行比较。微流控平台的商业化可以通过提供可靠的、具有成本效益的仪器来改善公共健康，该仪器可用于临床环境中的个性化癌症诊断。该技术通过微制造的可扩展性降低自动化细胞分析的成本，并通过减少样品体积实现对少量患者组织的多重测定，从而克服了当前的限制。类似的平台也可以适用于癌细胞生物学中的分子筛选和改进的高通量药物发现。

DESCRIPTION (provided by applicant): The main objective of this proposal is to develop a microfluidic platform for cancer drug toxicity screening in cultured human cells. While it is believed that improved information on a patient's individual cancer signature can aid diagnosis and treatment, the technology available to validate this claim is currently limiting. The long term goal of this work is to commercialize a microfluidic screening platform to provide a compact, low cost, automated screening system that can be used in the clinical setting. The specific aims of this proposal are to automate a previously developed microfluidic cell culture array and to demonstrate the feasibility and reproducibility of cancer drug toxicity screening in the microfluidic format. The design and fabrication of the addressable 8x3 unit microfluidic array will leverage expertise developed within the company related to soft lithography technology. Automation of fluidic delivery through the array will be accomplished through implementation of novel microfluidic valves controlled with an industrial pneumatic interface. Initial demonstration of cancer cell cytotoxicity will be collected on HeLa cells over 7 days exposure to anticancer drugs such as etoposide. Cell viability as well as apoptosis kinetics (quantified by fluorescence assay) will be collected in the array and experimental robustness determined. Response and statistical uniformity will be compared to the same assay performed in a 96-well plate. The commercialization of the microfluidic platform can improve public health by providing a reliable, cost effective instrument that can be used for personalized cancer diagnosis in the clinical setting. This technology overcomes current limitations by reducing the cost of automated cell analysis through the scalability of microfabrication, and by enabling multiplexed assays on a small amount of patient tissue through reduced sample volume. A similar platform can also be adapted for molecular screening in cancer cell biology and for improved high throughput drug discovery.