A Novel Combination of Thermoresponsive and Nanofibrillar Surface for Cell Cultur

描述（由申请人提供）：该SBIR项目旨在开发热响应纳米纤维作为细胞培养的微载体。纳米纤维表面为哺乳动物细胞的生长和功能提供了更像体内的生长表面。与胰蛋白酶或其他消化酶相比，热响应聚合物的使用允许细胞以温和、非侵入性且不那么麻烦的方式从基质释放。这种细胞分离方法消除了细胞培养过程中动物产品的使用，这是制药工艺的主要问题。该第二阶段项目描述了从纳米纤维和温度响应材料合成和制造微载体，用于大规模培养依赖于锚定的动物细胞。此外，还描述了磁性热响应微载体以帮助从培养系统中分离细胞。第二阶段计划的具体目标包括：1）将温敏纳米纤维制成微载体盘; 2）通过评估三种不同细胞类型的生长情况，优化温敏微载体在细胞培养中的使用; 3）将温敏纳米纤维制成微载体盘; 4）将温敏纳米纤维制成微载体盘; 5）将温敏纳米纤维制成微载体盘。3）比较和评估热响应微载体与广泛使用的市售微载体的效率; 4）合成磁性微载体并优化其在细胞和微载体悬浮液分离中的使用。 公共卫生相关性：使用动物细胞的基础研究和工业生物制药生产工艺具有许多安全和技术要求，例如不含动物来源物质的确定培养基、标准化、高产品产量、高产品浓度、放大潜力等。基于微载体的工艺满足许多这些要求。在微载体培养中，细胞生长在小球的表面上，或者在多孔结构的孔中作为多层生长，所述多孔结构通常通过轻轻搅拌悬浮在培养基中。微载体提供高得多的表面积与体积比，因此它们已被广泛用于扩增各种类型的粘附细胞，用于扩增细胞以大规模生产生长因子、疫苗和抗体。微载体的使用也在干细胞研究中获得认可，用于产生用于细胞治疗应用的大量细胞。细胞培养行业现在是每年300亿美元的生物制药市场的重要基础。随着疫苗、单克隆抗体、血栓溶解剂、干扰素、血液因子、重组蛋白和治疗酶的使用增加，使用哺乳动物细胞培养作为生产这些生物药物的主要方法的市场规模也将增长。

DESCRIPTION (provided by applicant): This SBIR project is designed to develop thermo-responsive nanofibers as microcarriers for cell culture. Nanofiber surfaces provide a more in vivo like growth surface for mammalian cell growth and function. The use of thermo-responsive polymer allows the cells to be released from the substrate in a gentle, non-invasive, and less cumbersome fashion compared to trypsin or other digestive enzymes. This method of cell detachment eliminates the use of animal products in the cell culture process, which is a major cause of concern for pharmaceutical manufacturing processes. This Phase II project describes the synthesis and fabrication of microcarriers from nanofibrillar and thermo-responsive materials for the large scale cultivation of anchorage dependent animal cells. Furthermore, a magnetic thermo-responsive microcarrier is also described to aid in the separation of cells from the culture system. This phase II project is expected to make the use of microcarrier culture more efficient for large scale production of vaccines, recombinant proteins, and cells for therapy, etc. Specific aims of this Phase II proposal include: 1) Fabricate thermo-responsive nanofibers into microcarrier discs; 2) Optimize the use of thermo-responsive microcarriers for cell culture by evaluating growth of three different cell types; 3) Compare and evaluate the efficiency of thermo-responsive microcarrier versus widely used commercially available microcarriers; 4) Synthesize magnetic microcarriers and optimize their use in cell and microcarrier suspension separation. PUBLIC HEALTH RELEVANCE: Basic research and industrial biopharmaceutical production processes using animal cells have a number of safety and technical requirements, such as defined media devoid of substances from animal origins, standardization, high product yield, high product concentration, scale-up potential, etc. Microcarrier based processes fulfill a number of these requirements. In microcarrier culture, cells grow either on the surface of small spheres or as multilayers in the pores of porous structures that are usually suspended in culture medium by gentle stirring. Microcarriers offer a much higher surface area to volume ratio and therefore they have been used extensively for amplifying various types of adherent cells for the expansion of cells for large scale production of growth factors, vaccines, and antibodies. The use of microcarriers is also gaining acceptance in the stem cell research for generating large quantities of cells for cell therapy applications. Cell culture industry is now a significant foundation of the $30 billion annual biopharmaceutical market. As the use of vaccines, monoclonal antibodies, thrombolytics, interferons, blood factors, recombinant proteins and therapeutic enzymes increases, the size of the market that uses mammalian cell culture as the primary method of producing these biopharmaceuticals will also grow.