Prometheus, Division of Skeletal Tissue Engineering, Katholieke Universiteit Leuven, Onderwijs & Navorsing 1, Herestraat 49, Box 813, 3000, Leuven, Belgium.
Biomechanics Research Unit, GIGA-In Sillico Medicine, University of Liège, B34 Quartier Hôpital, Avenue de l'Hôpital 1, 4000, Liège, Belgium.
Biotechnol J. 2019 Jul;14(7):e1800545. doi: 10.1002/biot.201800545. Epub 2019 May 15.
Bioreactors are crucial tools for the manufacturing of living cell-based tissue engineered products. However, to reach the market successfully, higher degrees of automation, as well as a decreased footprint still need to be reached. In this study, the use of a benchtop bioreactor for in vitro perfusion culture of scaffold-based tissue engineering constructs is assessed. A low-footprint benchtop bioreactor system is designed, comprising a single-use fluidic components and a bioreactor housing. The bioreactor is operated using an in-house developed program and the culture environment is monitored by specifically designed sensor ports. A gas-exchange module is incorporated allowing for heat and mass transfers. Titanium-based scaffolds are seeded with human periosteum-derived cells and cultured up to 3 weeks. The benchtop bioreactor constructs are compared to benchmark perfusion systems. Live/Dead stainings, DNA quantifications, glucose consumption, and lactate production assays confirm that the constructs cultured in the benchtop bioreactor grew similarly to the benchmark systems. Manual regulation of the system set points enabled efficient alteration of the culture environment in terms of temperature, pH, and dissolved oxygen. This study provides the necessary basis for the development of low-footprint, automated, benchtop perfusion bioreactors and enables the implementation of active environment control.
生物反应器是制造基于活细胞的组织工程产品的关键工具。然而，为了成功推向市场，仍需要提高自动化程度并减小占地面积。本研究评估了台式生物反应器在基于支架的组织工程构建物体外灌注培养中的应用。设计了一种低占地面积的台式生物反应器系统，包括一次性使用的流体组件和生物反应器外壳。该生物反应器使用内部开发的程序进行操作，并通过专门设计的传感器端口监测培养环境。结合了气体交换模块，允许进行热和质量传递。将钛基支架接种人骨膜来源细胞并培养长达 3 周。将台式生物反应器构建体与基准灌注系统进行比较。活/死染色、DNA 定量、葡萄糖消耗和乳酸生产测定证实，在台式生物反应器中培养的构建体与基准系统的生长情况相似。通过手动调节系统设定点，可以有效地改变温度、pH 值和溶解氧等培养环境。本研究为开发低占地面积、自动化的台式灌注生物反应器提供了必要的基础，并实现了主动环境控制。
