Microfluidic Cell Separation for Tissue Engineering and Regenerative Medicine

描述（由申请人提供）：本提案的主要目标是创建微流体细胞分离系统，以分离或丰富组织工程和再生医学的关键细胞类型。在传统的组织工程中，功能细胞类型必须在植入支架之前进行富集。在以细胞为基础的组织修复和再生方法中，驻留在不同组织类型中的干细胞和祖细胞必须在使用之前进行分离和表征。用于这些应用的微流体细胞分离系统的设计和制造是由以下观察引起的。首先，在功能细胞富集方面，微流控技术比预镀和使用细胞过滤器等最先进的方法更系统化。其次，微流体系统可以处理少量（微升量级）的样品，从而从少量供体组织中有效分离细胞。这些系统可以与体外细胞培养设备结合使用，此外，它们成本低，易于在临床环境中现场操作。第三，PI实验室最近的工作已经证明了微流体装置根据大小和亲和力分离细胞亚群的能力。本提案将著重于建立组织工程应用于四个领域的微流控细胞分离技术：心脏组织、皮肤组织、胃肠组织和血管组织。拟议的工作将与组织工程方面的专家合作进行。Milica radicic（心脏），Rebecca Carrier（肠道），Virna Sales & John Mayer（血管），Yaakov Nahmias & Martin Yarmush（皮肤/烧伤）。在为期3年的项目期间，将沿着独立的轨道实现以下目标：(1)设计和制造基于尺寸和粘附的微流体分离装置，以分离心脏和肠道组织中的细胞群；(2)设计一种基于粘附的微流体分离方法，通过正选择分离内皮祖细胞和皮肤干细胞，用于再生应用；(3)设计基于黏附的微流体分离方法，通过负选择分离心脏祖细胞和肠道干细胞。

DESCRIPTION (provided by applicant): The major goal of this proposal is to create microfluidic cell separation systems to isolate or enrich key cell types for tissue engineering and regenerative medicine. In conventional tissue engineering, functional cell types must be enriched prior to seeding onto scaffolds. In cell-based approaches to tissue repair and regeneration, stem and progenitor cells resident in different tissue types must be isolated and characterized prior to their use. The design and fabrication of microfluidic cell separation systems for these applications is motivated by the following observations. First, in functional cell enrichment, microfluidic techniques are more systematic compared to state of the art methods such as pre-plating and using cell strainers. Second, microfluidic systems can handle small (microliter-order) sample volumes, enabling effective cell separation from small quantities of donor tissue. These systems can be incorporated with in-vitro cell culture equipment and furthermore, they are low-cost and easily operated on-site in clinical settings. Third, recent work in the PI's laboratory has demonstrated the ability of microfluidic devices to separate cell subpopulations based on size and affinity. This proposal will focus on the creation of microfluidic cell separation technologies for tissue engineering applications in four areas: cardiac tissue, skin, gastrointestinal tissue, and vascular tissue. The proposed work will be carried out in collaboration with experts in tissue engineering: Drs. Milica Radisic (cardiac), Rebecca Carrier (intestinal), Virna Sales & John Mayer (vascular), and Yaakov Nahmias & Martin Yarmush (skin/burns). During the 3-year project period, the following aims will be pursued along independent tracks: (1) design and fabricate size- and adhesion-based microfluidic separation devices to separate cell populations in cardiac and intestinal tissue; (2) design an adhesion-based microfluidic separation approach to isolate endothelial progenitor cells and skin stem cells by positive selection for regenerative applications; and (3) design an adhesion-based microfluidic separation approach to isolate cardiac progenitor cells and intestinal stem cells by negative selection. PUBLIC HEALTH RELEVANCE: The ability to grow sections of certain tissue types from a small sample of donor tissue, tissue engineering has grown tremendously in recent years. Recent work has also demonstrated how stem and progenitor cells resident in adult tissue can play a role in repairing tissue. A key component of either of these approaches is the requirement to isolate or harvest certain cell types prior to utilizing them for repair and regenerative applications and this proposal aims to create novel tools for this purpose.