Microfluidic Reconstituted Mammary Tissue System

微流控重建乳腺组织系统

• 批准号: 6883038
• 负责人: Robert G Lowery
• 金额: $ 25.27万
• 依托单位: BELLBROOK LABS, LLC
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2006-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6883038
• 关键词: 3T3 cells; antineoplastics; biological signal transduction; biomimetics; biotechnology; breast neoplasms; cell cell interaction; cellular polarity; confocal scanning microscopy; controlled environment; drug screening /evaluation; extracellular matrix; fluorescence microscopy; fluorescent dye /probe; growth factor; immunocytochemistry; mammary epithelium; microfluidics; mixed tissue /cell culture; neoplasm /cancer chemotherapy; paracrine; steroid hormone; stromal cells; technology /technique development

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.

描述（由申请人提供）：发现乳腺癌的改良疗法需要准确重现完整乳腺组织细胞信号传导的测定系统。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体装置，用于在相邻的、可单独寻址的隔室中共培养人乳腺上皮细胞和基质细胞，这些隔室可以通过显微镜直接观察。该测定系统将能够在专门设计用于模拟乳腺肿瘤发生的体内组织背景的受控微环境中鉴定和验证分子药物靶标。 已经开发了使用包埋在凝胶基质中的细胞的三维器官型和异型培养方法来模拟乳腺中的ECM效应和旁分泌信号传导，但是它们对传统细胞培养系统（例如组织培养皿）的依赖限制了它们的准确性和实用性。在第一阶段，我们将制造一个专门设计用于重建和监测乳腺组织结构和功能的微流体装置。现有的用于在基质细胞附近3D培养上皮类器官的方法将针对该装置进行优化，以产生微流体重构乳腺组织系统（uF-rMTS）。uF-rMTS的规模和流体控制特性将提供比当前组织培养系统的本体水性环境更天然的微环境，并允许更好地复制体内信号传导现象。 在第二阶段，微流体装置将被多路复用以增加吞吐量，将纳入额外的正常和恶性细胞系，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。在完整的乳腺组织中开发一种探测致瘤信号传导过程的系统将加速发现更有效的乳腺癌治疗方法。