Microfluidic Cell Separation for Tissue Engineering and Regenerative Medicine

描述（由申请人提供）：该提案的主要目标是创建微流体细胞分离系统，以分离或富集用于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，在接种到支架上之前必须富集功能性细胞类型。在基于细胞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其使用之前被分离和表征。用于这些应用的微流控细胞分离系统的设计和制造是由以下观察所激发的。首先，在功能性细胞富集中，微流体技术与现有技术的方法如预接种和使用细胞过滤器相比更系统。其次，微流控系统可以处理小（微升量级）样品体积，从而能够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，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控设备基于大小和亲和力分离细胞亚群的能力。该提案将集中于创建用于组织工程应用的微流体细胞分离技术，用于四个领域：心脏组织，皮肤，胃肠道组织和血管组织。拟议的工作将与组织工程专家合作进行：Milica Radisic博士（心脏），Rebecca Carrier博士（肠道），Virna Sales和John Mayer博士（血管），以及Yaakov Nahovi和Martin Yarmush博士（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控分离装置，用于分离心脏和肠道组织中的细胞群;（2）设计基于粘附的微流控分离方法，用于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）设计一种基于粘附的微流控分离方法，通过负选择分离心肌祖细胞和肠干细胞。 公共卫生相关性：组织工程是从少量供体组织中培养出特定组织类型切片的能力，近年来发展迅速。最近的工作也证明了成体组织中的干细胞和祖细胞如何在修复组织中发挥作用。这两种方法的一个关键组成部分是需要在将某些细胞类型用于修复和再生应用之前分离或收获它们，该提案旨在为此目的创造新的工具。

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.