A Novel Combination of Thermoresponsive and Nanofibrillar Surface for Cell Cultur

描述（由申请人提供）：本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接近活体的生长表面。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一种温和、非侵入性和不那么麻烦的方式从底物中释放出来。这种细胞分离方法消除了在细胞培养过程中使用动物产品，这是药物制造过程中关注的主要原因。这个第二阶段的项目描述了纳米纤维和热响应材料的微载体的合成和制造，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。该二期项目预计将使微载体培养更有效地用于大规模生产疫苗、重组蛋白和用于治疗的细胞等。该第二阶段提案的具体目标包括：1)将热响应纳米纤维制成微载流子圆盘；2)通过对三种不同类型细胞的生长情况进行评价，优化热响应性微载体在细胞培养中的应用；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，优化其在细胞和微载体悬浮分离中的应用。

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.