Microfluidic Reconstituted Mammary Tissue System

描述(由申请人提供)：发现乳腺癌的改进治疗方法将需要准确概括完整乳腺组织细胞信号的检测系统。为了在体外忠实地重建乳腺组织结构和功能，我们将开发一种微流控设备，用于在相邻的、可单独寻址的隔间中共同培养人乳腺上皮细胞和基质细胞，这些隔间可以通过显微镜直接观察。这一检测系统将能够在受控微环境中识别和验证分子药物靶标，该微环境专门设计用于模拟乳腺肿瘤发生的体内组织背景。 利用植入凝胶基质中的细胞的三维器型和异型培养方法已被开发用于模拟乳房中的ECM效应和旁分泌信号，但它们对传统细胞培养系统(如组织培养皿)的依赖限制了它们的准确性和实用性。在第一阶段，我们将制造一种专门用于重建和监测乳腺组织结构和功能的微流控设备。现有的3D培养基质细胞附近上皮器官的方法将针对该设备进行优化，以产生微流控重组乳腺组织系统(UF-RMTs)。UF-RMT的规模和流体控制特性将提供比当前组织培养系统的整体水环境更自然的微环境，并允许更好地复制体内信号现象。 在第二阶段，微流控设备将被多路传输以增加吞吐量，更多的正常和恶性细胞株将被纳入，并将开发通过生长因子和类固醇激素探测旁分泌信号的方法。在完整的乳腺组织背景下探索致癌信号过程的系统的开发将加速发现更有效的乳腺癌治疗方法。

DESCRIPTION (provided by applicant): Discovery of improved therapies for breast cancer will require assay systems that accurately recapitulate the cellular signaling of intact mammary tissue. To reconstitute mammary tissue structure and function faithfully in vitro, we will develop a microfluidic device for co-culturing human mammary epithelial and stromal cells in adjacent, separately addressable compartments that can be directly observed by microscopy. This assay system will enable identification and validation of molecular drug targets in a controlled microenvironment specifically designed to mimic the in vivo tissue context of mammary tumorigenesis. Three dimensional organotypic and heterotypic culture methods using cells imbedded in gel matrices have been developed to model ECM effects and paracrine signaling in the breast, but their reliance on traditional cell culture systems (e.g. tissue culture dishes) limits their accuracy and utility. In Phase I, we will fabricate a microfluidic device designed specifically for reconstructing and monitoring mammary tissue structure and function. Existing methods for 3D culture of epithelial organoids in proximity to stromal cells will be optimized for the device to produce a Microfluidic Reconstituted Mammary Tissue System (uF-rMTS). The scale and fluid control properties of the uF-rMTS will provide a more native microenvironment than the bulk aqueous environment of current tissue culture systems and allow better replication of in vivo signaling phenomena. In Phase II, the microfluidics device will be multiplexed to increase throughput, additional normal and malignant cell lines will be incorporated, and methods will be developed to probe paracrine signaling by growth factors and steroid hormones. Development of a system for probing tumorigenic signaling processes in the context of intact mammary tissue will accelerate the discovery of more effective therapies for breast cancer.