Scinus Cell Expansion BV, Professor Bronkhorstlaan 10, Building 48, 3723 MB, Bilthoven, The Netherlands.
Leiden University Medical Centre, Albinusdreef 2, 2333 ZA, Leiden, The Netherlands.
J Transl Med. 2019 Jul 24;17(1):241. doi: 10.1186/s12967-019-1989-x.
Cell-based therapies have the potential to become treatment options for many diseases, but efficient scale-out of these therapies has proven to be a major hurdle. Bioreactors can be used to overcome this hurdle, but changing the culture method can introduce unwanted changes to the cell product. Therefore, it is important to establish parity between products generated using traditional methods versus those generated using a bioreactor.
Mesenchymal stromal cells (MSCs) are cultured in parallel using either traditional culture flasks, spinner vessels or a new bioreactor system. To investigate parity between the cells obtained from different methods, harvested cells are compared in terms of yield, phenotype and functionality.
Bioreactor-based expansion yielded high cell numbers (222-510 million cells). Highest cell expansion was observed upon culture in flasks [average 5.0 population doublings (PDL)], followed by bioreactor (4.0 PDL) and spinner flasks (3.3 PDL). Flow cytometry confirmed MSC identity (CD73, CD90 and CD105) and lack of contaminating hematopoietic cell populations. Cultured MSCs did not display genetic aberrations and no difference in differentiation and immunomodulatory capacity was observed between culture conditions. The response to IFNγ stimulation was similar for cells obtained from all culture conditions, as was the capacity to inhibit T cell proliferation.
The new bioreactor technology can be used to culture large amounts of cells with characteristics equivalent to those cultured using traditional, flask based, methods.
细胞疗法有可能成为许多疾病的治疗选择，但这些疗法的高效规模化生产已被证明是一个主要障碍。生物反应器可用于克服这一障碍，但改变培养方法可能会对细胞产品引入不必要的变化。因此，建立使用传统方法与使用生物反应器生产的产品之间的等效性非常重要。
间充质基质细胞（MSCs）分别使用传统培养瓶、搅拌瓶或新型生物反应器系统进行平行培养。为了研究不同方法获得的细胞之间的等效性，从不同方法获得的收获细胞在产量、表型和功能方面进行了比较。
基于生物反应器的扩增可获得大量细胞（2.22 亿至 5.1 亿个细胞）。在培养瓶中观察到最高的细胞扩增（平均 5.0 个倍增），其次是生物反应器（4.0 个倍增）和搅拌瓶（3.3 个倍增）。流式细胞术证实了 MSC 身份（CD73、CD90 和 CD105），并且不存在污染的造血细胞群体。培养的 MSC 未显示遗传异常，并且在分化和免疫调节能力方面，培养条件之间没有差异。来自所有培养条件的细胞对 IFNγ刺激的反应相似，抑制 T 细胞增殖的能力也相似。
新型生物反应器技术可用于培养大量具有与使用传统瓶式方法培养的细胞等效特征的细胞。
