Division of Cellular and Gene Therapies, Office of Tissue and Advanced Therapies, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, Maryland, USA.
Division of Cellular and Gene Therapies, Office of Tissue and Advanced Therapies, Center for Biologics Evaluation and Research, U.S. Food and Drug Administration, Silver Spring, Maryland, USA.
Cytotherapy. 2023 Sep;25(9):993-1005. doi: 10.1016/j.jcyt.2023.05.002. Epub 2023 May 29.
Human pluripotent stem cells (PSCs) hold a great promise for promoting regenerative medical therapies due to their ability to generate multiple mature cell types and for their high expansion potential. However, cell therapies require large numbers of cells to achieve desired therapeutic effects, and traditional two-dimensional static culture methods cannot meet the required production demand for cellular therapies. One solution to this problem is scaling up expansion of PSCs in bioreactors using culture strategies such as growing cells on microcarriers or as aggregates in suspension culture.
In this study, we directly compared PSC expansion and quality parameters in microcarrier- and aggregate-cultures grown in single-use vertical-wheel bioreactors.
We showed comparable expansion of cells on microcarriers and as aggregates by day 6 with a cell density reaching 2.2 × 10 cells/mL and 1.8 × 10 cells/mL and a fold-expansion of 22- and 18-fold, respectively. PSCs cultured on microcarriers and as aggregates were comparable with parallel two-dimensional cultures and with each other in terms of pluripotency marker expression and retention of other pluripotency characteristics as well as differentiation potential into three germ layers, neural precursor cells and cardiomyocytes.
Our study did not demonstrate a clear advantage between the two three-dimensional methods for the quality parameters assessed. This analysis adds support to the use of bioreactor systems for large scale expansion of PSCs, demonstrating that the cells retain key characteristics of PSCs and differentiation potential in suspension culture.
由于人类多能干细胞（PSCs）能够生成多种成熟细胞类型，并且具有较高的扩增潜力，因此在促进再生医学疗法方面具有巨大的潜力。然而，细胞疗法需要大量的细胞才能达到预期的治疗效果，而传统的二维静态培养方法无法满足细胞疗法所需的生产需求。解决这个问题的一种方法是使用培养策略（例如在微载体上生长细胞或在悬浮培养中作为聚集体生长）在生物反应器中放大 PSCs 的扩增。
在这项研究中，我们直接比较了在一次性垂直轮生物反应器中微载体和聚集体培养中 PSC 的扩增和质量参数。
我们显示，到第 6 天，微载体上和聚集体中的细胞扩增相当，细胞密度分别达到 2.2×10 个细胞/mL 和 1.8×10 个细胞/mL，扩增倍数分别为 22 倍和 18 倍。在微载体上和聚集体中培养的 PSCs 与平行二维培养物相似，并且在多能性标志物表达和保留其他多能性特征以及分化为三个胚层、神经前体细胞和心肌细胞的潜力方面彼此相似。
我们的研究没有显示出两种三维方法在评估的质量参数方面有明显的优势。这项分析支持使用生物反应器系统大规模扩增 PSCs，证明细胞在悬浮培养中保留了 PSCs 的关键特征和分化潜力。
