CAREER: Rolled Scaffold for High-Density Adherent Culture of Mammalian Cells

非技术摘要本项目由材料研究部生物材料计划资助，旨在开发一种名为滚动支架(RS)的新材料，并建立基于RS的下一代生物反应器，用于大规模细胞培养。随着生物技术的最新进展，动物细胞被用来生产基于蛋白质的药物和诊断抗体。此外，干细胞被研究为治疗以前无法治愈的退行性疾病的有效疗法。这些应用需要比目前可用的更高效率和可靠性的大型生物反应器。RS独特的微观结构提供了非常高的表面积与体积比，并允许培养介质在其中高效流动，使RS成为高密度贴壁培养的理想材料。与传统的生物反应器不同，基于RS的生物反应器将细胞与培养基库分开，允许充分的曝气，并在不损害细胞的情况下去除细胞生物废物。这些改进有望显著降低生物制药行业的生产成本，并促进干细胞在临床上的令人兴奋的应用。技术摘要搅拌槽式生物反应器悬浮培养被广泛应用于动物细胞的大规模培养。然而，有害的流体动剪切力和营养物质和气体传质速率不足限制了悬浮培养的效率和可靠性。在这个项目中，PI的目标是开发新型RS生物反应器，以克服悬浮培养的固有局限性。RS是一种带有间隔物的薄聚合物薄膜，这些间隔物被卷成一个圆柱体，其中包含许多相同的微流控通道。细胞在RS的内表面生长，培养液以层流的形式流经微流控通道。RS可以提供非常大的细胞培养面积和很高的营养物质和气体传质速率，剪切力最小。此外，细胞与介质储存库的分离使得基于RS的生物反应器具有新的配置。PI计划(1)开发RS的量产装置，(2)开发一种透析RS-生物反应器，可以有效地从介质中去除细胞废物，以高效地生产治疗性蛋白质，以及(3)实现一种具有低温介质存储的RS-生物反应器，可以延长热敏信号分子在培养液中的寿命，以用于负担得起的干细胞生产。此外，该项目旨在为高中开发自我可持续的STEM课程，通过各种外展活动在年轻学生中促进STEM素养，并通过积极参与研究将毕业生和本科生培养为生物材料领域的未来劳动力，优先考虑STEM中的女性和代表性较低的群体。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Nontechnical AbstractThis project supported by the Biomaterials program in the Materials Research Division is to develop a novel material called Rolled Scaffold (RS) and to build a next generation of bioreactors based on RS for large-scale cell culture. With recent progress in biotechnology, animal cells are used to produce protein-based drugs and diagnostic antibodies. Also, stem cells are investigated as effective cures for degenerative diseases that are previously incurable. These applications require large-scale bioreactors with higher efficiency and reliability than currently available ones. The unique microstructure of RS provides very high surface-area to volume ratio and allows efficient flow of culture media through it, making RS ideal material for high-density adherent culture. Unlike conventional bioreactors, the bioreactor based on RS separates the cells from the culture media reservoir, allowing sufficient aeration of media and removal of cellular bio-waste without cellular damages. These improvements are expected to significantly reduce the production cost in biopharmaceutical industries and facilitate the introduction of exciting clinical applications of stem cells. Technical AbstractSuspension culture in stirred tank bioreactors are widely used to culture animal cells in large scale. However, harmful hydrodynamic shear stress and insufficient mass transfer rate of nutrients and gases limit efficiency and reliability of suspension culture in large scale. In this project, PI aims to develop novel RS bioreactors to overcome inherent limitations of suspension culture. RS is a thin polymer film with spacers that are rolled into a cylinder containing numerous identical microfluidic channels. Cells are growing in inner surfaces of RS and culture media flows through the microfluidic channels in a laminar flow. The RS can provide very large cell culture area and high mass transfer rate of nutrients and gases with minimal shear stress. In addition, separation of cells from media reservoir allows novel configurations of RS-based bioreactors. PI plans (1) to develop a mass-production setup of RS, (2) to develop a dialysis RS-bioreactor that can effectively remove cellular waste from the media for efficient production of therapeutic proteins, and (3) to implement a RS-bioreactor with low-temperature media storage that can extend lifetime of heat-sensitive signaling molecules in culture media for affordable stem cell production. Besides, this project seeks to develop self-sustainable STEM curriculum for high school, to promote STEM literacy among young students through various outreach activities, and to train graduate and undergraduate student into future workforce in biomaterials area through active research participation with priorities to female and underrepresented groups in STEM.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.