Thomas Rob J, Anderson David, Chandra Amit, Smith Nigel M, Young Lorraine E, Williams David, Denning Chris
Loughborough University, UK.
Biotechnol Bioeng. 2009 Apr 15;102(6):1636-44. doi: 10.1002/bit.22187.
Large-scale manufacture of human embryonic stem cells (hESCs) is prerequisite to their widespread use in biomedical applications. However, current hESC culture strategies are labor-intensive and employ highly variable processes, presenting challenges for scaled production and commercial development. Here we demonstrate that passaging of the hESC lines, HUES7, and NOTT1, with trypsin in feeder-free conditions, is compatible with complete automation on the CompacT SelecT, a commercially available and industrially relevant robotic platform. Pluripotency was successfully retained, as evidenced by consistent proliferation during serial passage, expression of stem cell markers (OCT4, NANOG, TRA1-81, and SSEA-4), stable karyotype, and multi-germlayer differentiation in vitro, including to pharmacologically responsive cardiomyocytes. Automation of hESC culture will expedite cell-use in clinical, scientific, and industrial applications.
大规模制造人类胚胎干细胞（hESC）是其在生物医学应用中广泛使用的先决条件。然而，目前的hESC培养策略劳动强度大，且采用高度可变的流程，这给规模化生产和商业开发带来了挑战。在这里，我们证明在无饲养层条件下用胰蛋白酶传代hESC系HUES7和NOTT1，与在CompacT SelecT（一个商用且与工业相关的机器人平台）上完全自动化兼容。多能性得以成功保留，连续传代过程中的持续增殖、干细胞标志物（OCT4、NANOG、TRA1-81和SSEA-4）的表达、稳定的核型以及体外多胚层分化（包括分化为对药物有反应的心肌细胞）都证明了这一点。hESC培养的自动化将加快细胞在临床、科学和工业应用中的使用。
