Automated,scalable culture of human embryonic stem cells in feeder‐free conditions |
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| Authors: | Rob J. Thomas David Anderson Amit Chandra Nigel M. Smith Lorraine E. Young David Williams Chris Denning |
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| Affiliation: | 1. Healthcare Engineering Group, Wolfson School of Mechanical and Manufacturing Engineering, Loughborough University, Loughborough LE11 3TU, UK, telephone: +44‐1509‐22‐7601;2. fax: +44‐1509‐22‐7615;3. Wolfson Centre for Stem Cells, Tissue Engineering & Modelling (STEM), Centre for Biomolecular Sciences, University of Nottingham, Nottingham NG7 2RD, UK, telephone +44‐115‐82‐31236;4. Fax: +44‐115‐82‐31230;5. Department of Cytogenetics, Centre for Medical Genetics, Nottingham City Hospital NHS Trust, Nottingham, UK |
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| Abstract: | 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. Biotechnol. Bioeng. 2009;102: 1636–1644. © 2008 Wiley Periodicals, Inc. |
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| Keywords: | human embryonic stem cells automation CompacT SelecT scalability process control |
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