Frank Nathan D, Zylberberg Ezequiel, Roufai Mahajoub Bello, Gibb Stuart L, Miller Mindy M
Terumo Blood and Cell Technologies, Inc., Lakewood, Colorado, USA.
Akron Bio, Boca Raton, Florida, USA.
Cytotherapy. 2025 Mar;27(3):391-399. doi: 10.1016/j.jcyt.2024.11.011. Epub 2024 Dec 7.
The need for large-scale production of mesenchymal stromal cell (MSC)-based cellular therapeutics continues to grow around the globe. Manual cell expansion processes can be highly variable between operators, require significant hands-on time from skilled staff and, because of the large number of open manipulation steps required to produce cells in dose-relevant quantities, be prone to greater risk of contamination relative to automated processes. All of these can increase overall production costs and risks to the patient. In order to meet the needs of this growing industry, viable options for large-scale automation coupled with consistent and compliant ancillary materials needed to drive cell expansion are needed.
In the work described herein, the automated and functionally closed hollow-fiber bioreactor system Quantum Flex (Terumo Blood and Cell Technologies, Inc., Lakewood, CO, USA) was used in conjunction with Good Manufacturing Practice (GMP)-compliant, virus-inactivated human fibronectin (FN) from Akron Bio (Boca Raton, FL, USA) to expand MSCs to clinically relevant numbers. In order to assess the performance of Akron Bio's GMP-grade FN, use of this product in the production of MSCs was referenced against use of a research-use-only (RUO)-grade FN product used extensively for MSC expansion in Quantum. Because many MSC-based processes require passaging of cells to attain the appropriate number of cells needed, a two-passage process was employed comparing the transfer of MSCs expanded on RUO FN to RUO FN, GMP FN to GMP FN and RUO FN to GMP FN to assess the impacts of transitioning from one grade of FN to another, as a product might be required to do as it moves from pre-clinical to clinical stages and beyond.
No statistically significant differences were noted when MSCs were transferred from RUO FN to RUO FN, GMP FN to GMP FN or RUO FN to GMP FN in terms of harvest yield, population doubling time, seeding efficiency estimates or fold expansion. All MSCs harvested from all groups met International Society for Cell & Gene Therapy standards for MSCs in terms of protein marker expression measured by flow cytometry, adherence to plastic, downstream cell morphology and trilineage differentiation.
The combination of Quantum Flex as an expansion platform and Akron Bio's GMP FN is seen as an attractive option for larger-scale manufacture of GMP-grade MSC products.
全球对基于间充质基质细胞（MSC）的细胞疗法的大规模生产需求持续增长。手动细胞扩增过程在操作人员之间可能存在很大差异，需要熟练人员大量的实际操作时间，并且由于生产与剂量相关数量的细胞需要大量开放的操作步骤，相对于自动化过程更容易受到污染风险。所有这些都会增加总体生产成本和对患者的风险。为了满足这个不断发展的行业的需求，需要大规模自动化的可行选择以及驱动细胞扩增所需的一致且合规的辅助材料。
在本文所述的工作中，自动化且功能封闭的中空纤维生物反应器系统Quantum Flex（美国科罗拉多州莱克伍德的Terumo血液和细胞技术公司）与符合良好生产规范（GMP）、来自美国佛罗里达州博卡拉顿的Akron Bio公司的病毒灭活人纤连蛋白（FN）结合使用，将MSC扩增至临床相关数量。为了评估Akron Bio公司GMP级FN的性能，将该产品在MSC生产中的使用与在Quantum中广泛用于MSC扩增的仅用于研究（RUO）级FN产品的使用进行了比较。由于许多基于MSC的过程需要传代细胞以获得所需的适当细胞数量，因此采用了两步传代过程，比较在RUO FN上扩增的MSC向RUO FN、GMP FN向GMP FN以及RUO FN向GMP FN的转移，以评估从一种等级的FN转换到另一种等级的影响，因为产品在从临床前阶段到临床阶段及以后的过程中可能需要这样做。
当MSC从RUO FN转移到RUO FN、GMP FN转移到GMP FN或RUO FN转移到GMP FN时，在收获产量、群体倍增时间、接种效率估计或扩增倍数方面未观察到统计学上的显著差异。通过流式细胞术测量的蛋白质标志物表达、对塑料的粘附、下游细胞形态和三系分化方面，从所有组收获的所有MSC均符合国际细胞与基因治疗协会的MSC标准。
Quantum Flex作为扩增平台与Akron Bio公司的GMP FN的组合被视为大规模生产GMP级MSC产品的有吸引力的选择。
