A Novel Combination of Thermoresponsive and Nanofibrillar Surface for Cell Cultur

描述(申请人提供)：该SBIR项目旨在开发热响应性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。与胰酶或其他消化酶相比，使用温度响应性聚合物允许细胞以温和、非侵入性和不那么繁琐的方式从底物中释放出来。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，这是药物制造过程中令人担忧的主要原因。这个第二阶段的项目描述了由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锚定的动物细胞。此外，还描述了一种磁性温度响应微载体，以帮助细胞从培养系统中分离。这一二期项目有望使微载体培养更有效地用于大规模生产疫苗、重组蛋白和治疗用细胞等。这项二期计划的具体目标包括：1)将温度响应纳米纤维制备成微载体磁盘；2)通过评估三种不同类型的细胞的生长情况，优化温度响应微载体用于细胞培养的使用；3)比较和评估温度响应微载体与广泛使用的商用微载体的效率；4)合成磁性微载体并优化其在细胞和微载体悬浮分离中的使用。 与公众健康相关：使用动物细胞的基础研究和工业生物制药生产过程有许多安全和技术要求，例如不含动物来源物质的限定介质、标准化、高产品产量、高产品浓度、扩大潜力等。基于微载体的过程满足了许多这些要求。在微载体培养中，细胞要么生长在小球体的表面，要么以多层形式生长在多孔结构的孔隙中，这些多孔结构通常通过轻轻搅拌悬浮在培养介质中。微载体具有更高的表面积与体积比，因此被广泛用于放大各种类型的贴壁细胞，以扩大细胞规模生产生长因子、疫苗和抗体。微载体的使用在干细胞研究中也得到了认可，可以为细胞治疗应用产生大量细胞。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、单抗、溶栓剂、干扰素、血液因子、重组蛋白和治疗性酶的使用增加，将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扩大。

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.