Multi-functional particles for stem cell isolation and expansion

描述（由申请人提供）：本STTR I期提案的目标是开发新型多层微粒，并证明其分离和扩增干细胞用于医学研究和治疗应用的可行性。近年来，干细胞已被广泛应用于各种各样的应用，包括基于细胞的治疗，药物筛选和模型来了解疾病的发展。不幸的是，许多这些应用需要大量的干细胞，而这些干细胞是目前大多数细胞分离和培养方法无法产生的。现有方法的局限性包括使用有害的化学物质和酶进行细胞分离和扩增，以及在细胞传代过程中离心产生的强烈剪切力。为了克服这些缺点，我们将开发新的多层微颗粒，通过磁分离分离干细胞，使用温度敏感机制在370℃下结合细胞，通过3-D结构结构进行细胞扩增，通过温度敏感机制将细胞释放到室温下的表面。这些微粒将为干细胞的细胞分离、扩增和分离提供“一体式容器”，用于医疗应用，而无需使用有害化学品和机械力。对于第一阶段的研究，我们将制定和表征这些多层微粒（目标1）。然后将评估这些微粒在分离、扩增和分离干细胞（包括人血液中的内皮祖细胞（EPCs））方面的有效性（目标2）。该建议的成功完成可能为有效分离和扩增除干细胞外的各种细胞类型提供一种手段。从这项研究中获得的知识将大大提高我们对磁性细胞分离，干细胞扩增和细胞工程应用的微粒的理解。

DESCRIPTION (provided by applicant): The objective of this STTR Phase I proposal is to develop novel multi-layer microparticles and demonstrate their feasibility to isolate and expand stem cells for medical research and therapeutic applications. In recent years, stem cells have been used in a wide variety of applications, including cell- based therapies, drug screening, and models to understand the disease development. Unfortunately, many of these applications require large number of stem cells which cannot be produced by most of the current cell isolation and culture methods. The limitations of the current methods include the use of harmful chemicals and enzymes for cell isolation and expansion, and harsh shear forces from centrifugation during cell passaging process. To overcome these drawbacks, we will develop novel multi-layer microparticles that allow stem cell isolation via magnetic separation, cell attachment using a temperature-sensitive mechanism that binds cells at 370C, cell expansion via 3-D structural architecture, and cell detachment by a temperature-sensitive mechanism to release cells bound to the surface at room temperature. These microparticles will provide "all-in-one pot" for cell isolation, expansion, and detachment of stem cells for use in medical applications without the use of harmful chemicals and mechanical forces. For the phase I study, we will formulate and characterize these multilayer microparticles (Aim 1). The microparticles will then be evaluated for their effectiveness in isolation, expansion, and detachment of stem cells, including endothelial progenitor cells (EPCs) from human blood (Aim 2). The successful completion of this proposal may provide a means to effectively isolate and expand various cell types in addition to stem cells. The knowledge gained from this research should significantly improve our understanding of magnetic cell separation, stem cell expansion, and microparticles for cellular engineering applications.