Microfluidic Reconstituted Mammary Tissue System

描述（由申请人提供）：发现改进的乳腺癌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的结构和功能，我们将开发一种微流体装置，用于在相邻的、可单独定位的隔间中共同培养人类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。该检测系统将能够在一个受控的微环境中识别和验证分子药物靶点，该微环境专门设计用于模拟乳腺肿瘤发生的体内组织环境。

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.