Tirughana Revathiswari, Metz Marianne Z, Li Zhongqi, Hall Christine, Hsu David, Beltzer Jim, Annala Alexander J, Oganesyan Diana, Gutova Margarita, Aboody Karen S
Department of Developmental and Stem Cell Biology, Beckman Research Institute of City of Hope, Duarte, CA, USA.
Center for Biomedicine and Genetics, City of Hope, Duarte, CA, USA.
Mol Ther Methods Clin Dev. 2018 Jul 7;10:48-56. doi: 10.1016/j.omtm.2018.05.006. eCollection 2018 Sep 21.
Cell-based therapies hold great promise for a myriad of clinical applications. However, as these therapies move from phase I to phase II and III trials, there is a need to improve scale-up of adherent cells for the production of larger good manufacturing practice (GMP) cell banks. As we advanced our neural stem cell (NSC)-mediated gene therapy trials for glioma to include dose escalation and multiple treatment cycles, GMP production using cell factories (CellStacks) generated insufficient neural stem cell (NSC) yields. To increase yield, we developed an expansion method using the hollow fiber quantum cell expansion (QCE) system. Seeding of 5.2 × 10 NSCs in a single unit yielded up to 3 × 10 cells within 10 days. These QCE NSCs showed genetic and functional stability equivalent to those expanded by conventional flask-based methods. We then expanded the NSCs in 7 units simultaneously to generate a pooled GMP-grade NSC clinical lot of more than 1.5 × 10 cells in only 9 days versus 8 × 10 over 6 weeks in CellStacks. We also adenovirally transduced our NSCs within the QCE. We found the QCE system enabled rapid cell expansion and increased yield while maintaining cell properties and reducing process time, labor, and costs with improved efficiency and reproducibility.
基于细胞的疗法在众多临床应用中具有巨大潜力。然而，随着这些疗法从I期试验进入II期和III期试验，需要改进贴壁细胞的扩大培养，以生产更大规模的药品生产质量管理规范（GMP）细胞库。随着我们将神经干细胞（NSC）介导的胶质瘤基因治疗试验推进到包括剂量递增和多个治疗周期，使用细胞工厂（CellStacks）进行的GMP生产产生的神经干细胞（NSC）产量不足。为了提高产量，我们开发了一种使用中空纤维量子细胞扩增（QCE）系统的扩增方法。在单个单元中接种5.2×10个神经干细胞，在10天内可产生多达3×10个细胞。这些QCE神经干细胞显示出与通过传统基于培养瓶的方法扩增的细胞相当的遗传和功能稳定性。然后，我们同时在7个单元中扩增神经干细胞，仅用9天就产生了一批超过1.5×10个细胞的GMP级神经干细胞临床批次，而在CellStacks中则需要6周才能产生8×10个细胞。我们还在QCE内对神经干细胞进行腺病毒转导。我们发现QCE系统能够实现快速细胞扩增并提高产量，同时保持细胞特性，减少处理时间、劳动力和成本，提高效率和可重复性。
