Osiecki Michael J, Michl Thomas D, Kul Babur Betul, Kabiri Mahboubeh, Atkinson Kerry, Lott William B, Griesser Hans J, Doran Michael R
Institute of Health and Biomedical Innovation, Queensland University of Technology at the Translational Research Institute, Brisbane, Queensland, Australia.
School of Chemistry, Physics and Mechanical Engineering, Science and Engineering Faculty, Queensland University of Technology Brisbane, Queensland, Australia.
PLoS One. 2015 Dec 14;10(12):e0144941. doi: 10.1371/journal.pone.0144941. eCollection 2015.
Large numbers of Mesenchymal stem/stromal cells (MSCs) are required for clinical relevant doses to treat a number of diseases. To economically manufacture these MSCs, an automated bioreactor system will be required. Herein we describe the development of a scalable closed-system, packed bed bioreactor suitable for large-scale MSCs expansion. The packed bed was formed from fused polystyrene pellets that were air plasma treated to endow them with a surface chemistry similar to traditional tissue culture plastic. The packed bed was encased within a gas permeable shell to decouple the medium nutrient supply and gas exchange. This enabled a significant reduction in medium flow rates, thus reducing shear and even facilitating single pass medium exchange. The system was optimised in a small-scale bioreactor format (160 cm2) with murine-derived green fluorescent protein-expressing MSCs, and then scaled-up to a 2800 cm2 format. We demonstrated that placental derived MSCs could be isolated directly within the bioreactor and subsequently expanded. Our results demonstrate that the closed system large-scale packed bed bioreactor is an effective and scalable tool for large-scale isolation and expansion of MSCs.
治疗多种疾病所需的临床相关剂量需要大量间充质干/基质细胞（MSC）。为了经济地生产这些MSC，将需要一个自动化生物反应器系统。在此，我们描述了一种适用于大规模MSC扩增的可扩展封闭系统填充床生物反应器的开发。填充床由经空气等离子体处理的熔融聚苯乙烯颗粒形成，使其具有与传统组织培养塑料相似的表面化学性质。填充床被包裹在透气壳内，以使培养基营养供应和气体交换解耦。这使得培养基流速显著降低，从而减少剪切力，甚至便于单程培养基更换。该系统在小规模生物反应器形式（160 cm²）中用小鼠来源的表达绿色荧光蛋白的MSC进行了优化，然后扩大到2800 cm²的形式。我们证明胎盘来源的MSC可以直接在生物反应器内分离并随后扩增。我们的结果表明，封闭系统大规模填充床生物反应器是用于大规模分离和扩增MSC的有效且可扩展的工具。
