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支持，旨在开发一种称为卷式支架（RS）的新型材料，并基于RS构建下一代生物反应器，用于大规模细胞培养。随着生物技术的最新进展，动物细胞被用于生产基于蛋白质的药物和诊断抗体。此外，干细胞被研究为以前无法治愈的退行性疾病的有效治疗方法。这些应用需要比现有生物反应器具有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，并允许培养基有效地流过它，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储存器分离，允许培养基充分通气并在不损伤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，并促进干细胞令人兴奋的临床应用的引入。搅拌式生物反应器中的悬浮培养被广泛应用于动物细胞的大规模培养。然而，有害的流体动力学剪切应力和营养物质和气体的传质速率不足限制了大规模悬浮培养的效率和可靠性。在这个项目中，PI旨在开发新型RS生物反应器，以克服悬浮培养的固有局限性。RS是一种具有间隔物的薄聚合物膜，该间隔物被卷成包含许多相同的微流体通道的圆柱体。细胞在RS的内表面生长，并且培养基以层流流过微流体通道。RS可以提供非常大的细胞培养面积和营养物和气体的高传质速率以及最小的剪切应力。此外，从培养基储器分离细胞允许基于RS的生物反应器的新配置。PI计划（1）开发RS的大规模生产装置，（2）开发透析RS-生物反应器，可有效去除培养基中的细胞废物，以有效生产治疗性蛋白质，以及（3）实施具有低温培养基储存的RS-生物反应器，可延长培养基中热敏信号分子的寿命，以生产负担得起的干细胞。此外，该项目旨在为高中开发自我维持的STEM课程，通过各种外展活动促进年轻学生的STEM扫盲，并通过积极参与研究，将研究生和本科生培养成生物材料领域未来的劳动力，优先考虑STEM中的女性和代表性不足的群体。该奖项反映了NSF的法定使命，并通过使用基金会的学术价值和更广泛的影响审查标准。

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.