Pharmaceutical Production Research Facility, Schulich School of Engineering, University of Calgary, Calgary, Alberta, Canada.
Department of Biomedical Engineering, Schulich School of Engineering, University of Calgary, Calgary, Alberta, Canada.
Biotechnol Bioeng. 2024 Jan;121(1):192-205. doi: 10.1002/bit.28557. Epub 2023 Sep 29.
Equine mesenchymal stromal cells (MSCs) have been found to be beneficial for the treatment of many ailments, including orthopedic injuries, due to their superior differentiation potential and immunomodulating properties. Cell therapies require large cell numbers, which are not efficiently generated using conventional static expansion methods. Expansion of equine cord blood-derived MSCs (eCB-MSCs) in bioreactors, using microcarriers as an attachment surface, has the potential to generate large numbers of cells with increased reproducibility and homogeneity compared with static T-flask expansion. This study investigated the development of an expansion process using Vertical-Wheel (VW) bioreactors, a single-use bioreactor technology that incorporates a wheel instead of an impeller. Initially, microcarriers were screened at small scale to assess eCB-MSC attachment and growth and then in bioreactors to assess cell expansion and harvesting. The effect of different donors, serial passaging, and batch versus fed batch were all examined in 0.1 L VW bioreactors. The use of VW bioreactors with an appropriate microcarrier was shown to be able to produce cell densities of up to 1E6 cells/mL, while maintaining cell phenotype and functionality, thus demonstrating great potential for the use of these bioreactors to produce large cell numbers for cell therapies.
马间充质干细胞（MSCs）因其具有优异的分化潜能和免疫调节特性，已被发现对许多疾病（包括骨科损伤）的治疗有益。细胞疗法需要大量的细胞，而传统的静态扩增方法并不能有效地产生这些细胞。在生物反应器中使用微载体作为附着表面来扩增脐带血来源的马间充质干细胞（eCB-MSCs），与静态 T 瓶扩增相比，具有产生大量细胞的潜力，并且具有更高的重现性和均一性。本研究开发了一种使用垂直轮（VW）生物反应器的扩增工艺，这是一种使用轮而不是叶轮的一次性生物反应器技术。最初，在小范围内筛选微载体，以评估 eCB-MSC 的附着和生长，然后在生物反应器中评估细胞的扩增和收获。在 0.1L VW 生物反应器中，还检查了不同供体、传代和分批与补料分批的影响。结果表明，使用适当的微载体的 VW 生物反应器能够产生高达 1E6 个细胞/ml 的细胞密度，同时保持细胞表型和功能，因此，这些生物反应器在用于细胞疗法的大量细胞生产方面具有很大的潜力。
