Microfluidic Platform for Cancer Cell Culture and Analysis

描述(申请人提供)：这项第二阶段的SBIR建议开发一种用于癌细胞体外筛选的微流控细胞培养和分析平台。这项研究的最终结果将是提供一个功能齐全的系统(自动化仪器和一次性微流体)，既可以用于研究实验室，也可以用于制药实验室。这将得到验证，并应用于我们合作研究所正在进行的研究，目的是在一个由大约60个乳腺癌细胞株组成的小组中描绘Raf-MEK-ERK通路，以改进对治疗反应的预测。 与当前基于细胞的筛选技术(以96孔板为基础)相比，微流控平台将提供主要优势，包括：1)改进了小细胞样本的处理(每个阵列微升)，2)能够为表型分析设计更相关的微环境，3)实现多路连续流动实验，4)细胞培养自动化的时间和成本减少10倍-100倍。此外，该平台的设计使得特定于应用的微流控阵列可以与单个系统一起使用，从而增加了该技术的灵活性和影响力。 该项目的第一个主要目标将是设计一个自动化微流控筛选平台。主要任务是优化第一阶段原型的设计，扩大到384井格式，并改进控制系统以实现高通量操作。我们之前工作中开发的三项关键创新将进一步扩展以实现这一目标：1)微流控网络和灌流屏障的设计，以更好地模拟体内培养条件；2)使用气动压力驱动歧管进行多路、非湿式纳升体积的泵送；以及3)能够将微流控阵列格式化为SBS标准的制造工艺，使其与当前的96和384井机器人筛选仪器兼容。 第二个主要目标是将该系统应用于我们合作研究所的癌细胞筛查计划。这将涉及微流控技术可以提供现有工具所不能提供的有利好处的领域。具体的应用包括：1)基于流动的药物暴露，2)细胞侵袭分析，3)3D ECM培养，4)基质细胞的培养条件，5)与RNAi方法的整合。

DESCRIPTION (provided by applicant): This Phase II SBIR proposes to develop a microfluidic cell culture and analysis platform for the in vitro screening of cancer cells. The end result of this research will be to deliver a fully functional system (automated instrument and disposable microfluidics) that can be used in both research and pharmaceutical labs. This will be validated and applied to the research underway at our collaborating institute for the purpose of profiling the Raf-MEK-ERK pathway in a panel of ~60 breast cancer cell lines for improved prediction of therapeutic response. The microfluidic platform will provide key advantages over the current cell based screening technology (96-well plate based), including: 1) improved handling of small cell samples (micoliters per array), 2) the ability to design more relevant microenvironments for phenotype analysis, 3) enabling multiplexed continuous flow experimentation, and 4) 10X-100X reduction of time and cost for cell culture automation. In addition, the platform is designed such that application specific microfluidic arrays can be utilized with a single system, increasing the flexibility and impact of the technology. The first major aim of this project will be to engineer an automated microfluidic screening platform. The main tasks are to optimize the design of the Phase I prototype, scale-up to a 384 well format, and refine the control system for high throughput operation. Three key innovations developed in our previous work will be further expanded to complete this aim: 1) the design of microfluidic networks and perfusion barriers to better approximate in vivo culture conditions, 2) the use of a pneumatic pressure driven manifold for multiplexed, non-wetted pumping of nanoliter volumes, and 3) the fabrication process that enables formatting the microfluidic arrays to SBS standards, making it compatible with current 96 and 384 well robotic screening instrumentation. The second major aim will be to apply this system to the cancer cell screening program at our collaborating institute. This will address areas where microfluidic technology can offer enabling benefits not possible with existing tools. Specific applications include: 1) flow based drug exposure, 2) cell invasion assay, 3) 3D ECM culture, 4) medium conditioning by stromal cells, and 5) integration with RNAi methods.