Biomedical Engineering Graduate Program, University of Calgary, 2500 University Drive NW, Calgary, AB T2N 1N4, Canada.
Pharmaceutical Production Research Facility, University of Calgary, 2500 University Drive NW, Calgary, AB T2N 1N4, Canada.
Int J Mol Sci. 2023 Mar 8;24(6):5152. doi: 10.3390/ijms24065152.
Regenerative therapies for the treatment of peripheral nerve and spinal cord injuries can require hundreds of millions of autologous cells. Current treatments involve the harvest of Schwann cells (SCs) from nerves; however, this is an invasive procedure. Therefore, a promising alternative is using skin-derived Schwann cells (Sk-SCs), in which between 3-5 million cells can be harvested from a standard skin biopsy. However, traditional static planar culture is still inefficient at expanding cells to clinically relevant numbers. As a result, bioreactors can be used to develop reproducible bioprocesses for the large-scale expansion of therapeutic cells. Here, we present a proof-of-concept SC manufacturing bioprocess using rat Sk-SCs. With this integrated process, we were able to simulate a feasible bioprocess, taking into consideration the harvest and shipment of cells to a production facility, the generation of the final cell product, and the cryopreservation and shipment of cells back to the clinic and patient. This process started with 3 million cells and inoculated and expanded them to over 200 million cells in 6 days. Following the harvest and post-harvest cryopreservation and thaw, we were able to maintain 150 million viable cells that exhibited a characteristic Schwann cell phenotype throughout each step of the process. This process led to a 50-fold expansion, producing a clinically relevant number of cells in a 500 mL bioreactor in just 1 week, which is a dramatic improvement over current methods of expansion.
用于治疗周围神经和脊髓损伤的再生疗法可能需要数亿个自体细胞。目前的治疗方法涉及从神经中采集雪旺细胞（SCs）；然而，这是一种有创的程序。因此，一种很有前途的替代方法是使用皮肤衍生的雪旺细胞（Sk-SCs），从标准皮肤活检中可以采集到 300 万到 500 万个细胞。然而，传统的静态平面培养仍然不能有效地将细胞扩增到临床相关数量。因此，可以使用生物反应器来开发可重复的生物工艺，以大规模扩增治疗细胞。在这里，我们提出了一个使用大鼠 Sk-SCs 的 SC 制造生物工艺的概念验证。通过这个集成的过程，我们能够模拟一个可行的生物工艺，考虑到细胞的采集和运输到生产设施，最终细胞产品的生成，以及细胞的冷冻保存和运输回临床和患者。这个过程从 300 万个细胞开始，在 6 天内将其接种并扩增到 2 亿多个细胞。在收获和收获后冷冻保存和解冻后，我们能够保持 1.5 亿个存活细胞，这些细胞在整个过程的每一步都表现出典型的雪旺细胞表型。这一过程导致了 50 倍的扩增，在一个 500 毫升的生物反应器中仅用 1 周就产生了临床相关数量的细胞，这是对当前扩张方法的显著改进。
