SBIR Phase I: Novel Polymer Brush-like Architecture for Temperature Dependent Cell Release and Attachment

这个小企业创新研究（SBIR）第一阶段项目旨在提供两种独特的定制热响应表面，用于附着和释放弱和强粘附细胞。商业上可用的表面是高度交联的水凝胶，不能很好地与强或弱粘附细胞一起工作。目标是通过在培养表面提供新型聚合物刷状结构来改变当前的热响应细胞培养技术，以减少蛋白质吸附，并允许有效的水合作用释放强粘附细胞。此外，聚合物刷状结构的附加功能将增加弱粘附细胞表面的粘附强度。本提案的主要研究目标包括：1)合成用于强粘附细胞的光反应性热响应刷结构；2)合成用于弱粘附细胞的功能化刷结构；3)开发各种不同的培养皿涂层，目前由于使用的方法而无法在市场上获得。目标是在室温下30分钟内释放90%的强贴壁细胞，80%的弱贴壁细胞附着。该项目更广泛的影响/商业潜力将是产生功能性细胞或细胞片。生理活性功能细胞的产生往往是技术发展的障碍。例如，从传统生长表面回收的细胞通过消化酶和胰蛋白酶、EDTA等化学物质释放，从而改变信号转导途径，影响细胞的整体活力。使用聚n -异丙基丙烯酰胺（PNIPAAm）的热反应性细胞培养皿避免了消化酶的使用；然而，使用热反应性培养器皿的主要障碍仍然存在。市售热响应板适用于几种细胞系，但不适用于强粘附或弱粘附细胞类型，并且仅适用于多孔格式。第一阶段拨款将用于开发新一代的热响应细胞培养皿，以获得广泛的市场接受，并成为一项使能技术。该工艺使用简单的设备，易于制造，具有成本效益，并且可用于组织工程和高通量应用所需的格式。因此，以前认为不可能的细胞类型的临床应用可能会实现，细胞治疗、药物发现和组织工程应用可能会加快步伐。

This Small Business Innovation Research (SBIR) Phase I project is designed to offer two uniquely tailored thermoresponsive surfaces for the attachment and release of weakly and strongly adherent cells. The commercially available surfaces are highly crosslinked hydrogels that do not work well with either strongly or weakly adherent cells. The goal is to transform the current thermoresponsive cell culture technology by providing novel polymer brush-like architectures to the culture surface to reduce protein adsorption, and allow efficient hydration to release the strongly adherent cells. Moreover, addition of functionalities to polymer brush-like architectures will increase the adhesive strength of the surfaces for weakly adherent cells. Major research objectives of this proposal include: 1) Synthesis of photoreactive thermoresponsive brush architectures for strongly adherent cells 2) Synthesis of functionalized brush architectures for weakly adherent cells and 3) Develop coatings in a variety of different cultureware not presently available in market due to the method used. The goal is 90% release of strongly adherent cells within 30 minutes at room temperature and 80% attachment of weakly adherent cells. The broader impact /commercial potential of this project will be the generation of functional cells or cell sheets. The generation of physiologically active functional cells is often the roadblock in technology development. For example, cells recovered from conventional growth surfaces are released using digestive enzymes and chemicals such as Trypsin and EDTA, which alter signal transduction pathways and affect overall cell viability. Thermoresponsive cell cultureware involving the use of Poly N-isopropylacrylamide (PNIPAAm) obviates the use of digestive enzymes; however, major roadblocks with the use of thermoresponsive cultureware remain. The commercially available thermoresponsive plates work well with several cell lines, but do not work well with strongly adherent or weakly adherent cell types, and are available only in multiwell formats. This Phase I grant will develop a new generation of thermoresponsive cell cultureware that could gain wide market acceptance, and become an enabling technology. The process uses simple equipment, is manufacture-friendly, cost-effective, and is available in formats needed for tissue engineering and high-throughput applications. Thus, clinical applications of cell types that were previously considered impossible may be realized, and cell therapy, drug discovery, and tissue engineering applications may proceed at an accelerated pace.