Department of Bioengineering and iBB, Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Avenida Rovisco Pais, Lisboa, 1049-001, Portugal.
The Discoveries Center for Regenerative and Precision Medicine, Lisbon Campus, Instituto Superior Técnico, Universidade de Lisboa, Avenida Rovisco Pais, Lisboa, 1049-001, Portugal.
Biotechnol J. 2019 Aug;14(8):e1800716. doi: 10.1002/biot.201800716. Epub 2019 May 31.
Mesenchymal stromal cells (MSC) hold great promise for tissue engineering applications and cell-based therapies. Large cell doses (>1 × 10 cells kg ) and Good Manufacturing Practices (GMP)-compliant processes are however required for clinical purposes. Here, a serum- and xenogeneic-free (S/XF) microcarrier-based culture system is established for the expansion of human umbilical cord matrix (UCM)- and adipose tissue (AT)-derived MSC using the Vertical-Wheel system (PBS-0.1 MAG; PBS Biotech). UCM and AT MSC are expanded to maximum cell densities of 5.3 ± 0.4 × 10 cell mL (n = 3) and 3.6 ± 0.7 × 10 cell mL (n = 3), respectively, after 7 days of culture, while maintaining their identity, according to standard criteria. An economic evaluation of the process transfer from T-flasks to PBS-0.1 MAG shows a reduction in the costs associated with the production of a dose for an average 70 kg adult patient (i.e., 70 million cells). Costs decrease from $17.0 K to $11.1 K for UCM MSC and from $21.5 K to $11.1 K for AT MSC, proving that the transition to Vertical-Wheel reactors provides a cost-effective alternative for MSC expansion. The present work reports the establishment of a scalable and cost-effective culture platform for the manufacturing of UCM and AT MSC in a S/XF microcarrier-based system.
间充质基质细胞（MSC）在组织工程应用和基于细胞的治疗中具有巨大的应用前景。然而，出于临床应用的目的，需要大剂量的细胞（>1×10^6 细胞/kg）和符合良好生产规范（GMP）的工艺。在此，建立了一种无血清和无异种（S/XF）微载体培养系统，使用 Vertical-Wheel 系统（PBS-0.1 MAG；PBS Biotech）来扩增人脐带基质（UCM）和脂肪组织（AT）来源的 MSC。UCM 和 AT MSC 在培养 7 天后，分别可扩增至 5.3±0.4×10^6 细胞/mL（n=3）和 3.6±0.7×10^6 细胞/mL（n=3）的最大细胞密度，同时根据标准标准保持其特性。将工艺从 T 瓶转移到 PBS-0.1 MAG 的经济评估表明，与生产 70 公斤成年患者（即 7000 万细胞）剂量相关的成本降低。UCM MSC 的成本从$17.0 K 降低到$11.1 K，AT MSC 的成本从$21.5 K 降低到$11.1 K，证明向 Vertical-Wheel 反应器的转变为 MSC 扩增提供了一种具有成本效益的替代方案。本工作报道了在 S/XF 微载体系统中建立可扩展且具有成本效益的 UCM 和 AT MSC 制造的培养平台。
