Department of Biochemistry and Molecular Biology, Faculty of Medicine, University of Calgary, Calgary, Alberta, Canada.
Tissue Eng Part C Methods. 2010 Aug;16(4):573-82. doi: 10.1089/ten.TEC.2009.0228.
Since the derivation of human embryonic stem (hES) cells, their translation to clinical therapies has been met with several challenges, including the need for large-scale expansion and controlled differentiation processes. Suspension bioreactors are an effective alternative to static culture flasks as they enable the generation of clinically relevant cell numbers with greater efficacy in a controlled culture system. We, along with other groups, have developed bioreactor protocols for the expansion of pluripotent murine ES cells. Here we present a novel bioreactor protocol that yields a 25-fold expansion of hES cells over 6 days. Using immunofluorescence, flow cytometry, and teratoma formation assays, we demonstrated that these bioreactor cultures retained high levels of pluripotency and a normal karyotype. Importantly, the use of bioreactors enables the expansion of hES cells in the absence of feeder layers or matrices, which will facilitate the adaptation of good manufacturing process (GMP) standards to the development of hES cell therapies.
自从人类胚胎干细胞（hES）的衍生以来，其向临床治疗的转化遇到了几个挑战，包括需要大规模扩增和受控分化过程。悬浮生物反应器是静态培养瓶的有效替代方法，因为它们能够在受控培养系统中以更高的效率生成具有临床相关性的细胞数量。我们与其他小组一起，为多能鼠胚胎干细胞的扩增开发了生物反应器方案。在这里，我们提出了一种新的生物反应器方案，可在 6 天内使 hES 细胞扩增 25 倍。通过免疫荧光、流式细胞术和畸胎瘤形成分析，我们证明这些生物反应器培养物保持了高水平的多能性和正常的核型。重要的是，生物反应器的使用可以在没有饲养层或基质的情况下扩增 hES 细胞，这将有助于将良好生产规范（GMP）标准应用于 hES 细胞治疗的开发。
