Automated, Scalable Culture of Human Embryonic Stem Cells in Feeder-Free Conditions
Automated, Scalable Culture of Human Embryonic Stem Cells in Feeder-Free Conditions
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DOI：
10.1002/bit.22187
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发表时间：
2009-04-15
影响因子：
3.8
通讯作者：
Denning, Chris
中科院分区：
文献类型：
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作者：
Thomas, Rob J.;Anderson, David;Denning, Chris
人类胚胎干细胞（hESC）的大规模生产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培养策略是劳动密集型的，并且采用高度可变的工艺，这对规模化生产和商业化开发提出了挑战。在这里，我们证明了在无饲养条件下用胰蛋白酶传代hESC系HUES 7和NOTT 1与CompacT SelecT（一种市售和工业相关的机器人平台）上的完全自动化兼容。成功地保留了多能性，如通过连续传代期间的一致增殖、干细胞标志物（OCT 4、NANOG、TRA 1 -81和SSEA-4）的表达、稳定的核型和体外多胚层分化（包括向细胞周期反应性心肌细胞的分化）所证明的。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。
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.