Jorgenson Kristen D, Hart David A, Krawetz Roman, Sen Arindom
Pharmaceutical Production Research Facility (PPRF), Schulich School of Engineering, University of Calgary, 2500 University Drive NW, Calgary, AB, Canada T2N 1N4.
McCaig Institute for Bone and Joint Health, Cumming School of Medicine, University of Calgary, 3280 Hospital Drive NW, Calgary, AB, Canada T2N 4Z6.
Stem Cells Int. 2018 Mar 27;2018:8431053. doi: 10.1155/2018/8431053. eCollection 2018.
The chondrogenic potential of synovial fluid-derived mesenchymal stem cells (SF-MSCs) supports their use in cartilage regeneration strategies. However, their paucity in synovial fluid necessitates their proliferation in culture to generate clinically relevant quantities. Here it was determined that 125 mL stirred suspension bioreactors utilizing Cytodex-3 microcarrier beads represent a viable platform for the proliferation of these cells. During the inoculation phase, a bead loading of 2 g/L, an inoculation ratio of 4.5 cells/bead, and continuous agitation at 40 rpm in a medium with 5% serum resulted in high cell attachment efficiencies and a subsequent overall cell fold expansion of 5.7 over 8 days. During the subsequent growth phase, periodic addition of new microcarriers and fresh medium increased culture longevity, resulting in a 21.3 cell fold increase over 18 days in the same vessel without compromising the defining characteristics of the cells. Compared to static tissue culture flasks, a bioreactor-based bioprocess requires fewer handling steps, is more readily scalable, and for the same cell production level, has a lower operating cost as it uses approximately half the medium. Therefore, stirred suspension bioreactors incorporating microcarrier technology represent a viable and more efficient platform than tissue culture flasks for the generation of SF-MSCs in culture.
滑膜液源性间充质干细胞（SF-MSCs）的软骨生成潜能支持其在软骨再生策略中的应用。然而，滑膜液中这些细胞数量稀少，因此需要在培养中进行增殖以获得临床相关数量。研究确定，使用Cytodex-3微载体珠的125 mL搅拌悬浮生物反应器是这些细胞增殖的可行平台。在接种阶段，微载体珠负载量为2 g/L，接种比例为4.5个细胞/珠，在含5%血清的培养基中以40 rpm持续搅拌，可实现较高的细胞附着效率，随后在8天内细胞总体扩增倍数达5.7倍。在随后的生长阶段，定期添加新的微载体和新鲜培养基可延长培养时间，在同一容器中18天内细胞扩增倍数达21.3倍，且不影响细胞的特性。与静态组织培养瓶相比，基于生物反应器的生物工艺操作步骤更少，更易于扩大规模，并且在相同细胞产量水平下，由于使用的培养基约为一半，运行成本更低。因此，结合微载体技术的搅拌悬浮生物反应器是比组织培养瓶更可行、更高效的用于在培养中生成SF-MSCs的平台。
