2134/4240 Rob Thomas Rob Thomas David Anderson David Anderson Amit Chandra Amit Chandra Nigel M. Smith Nigel M. Smith Lorraine E. Young Lorraine E. Young David Williams David Williams Chris Denning Chris Denning Automated, scalable culture of human embryonic stem cells in feeder free conditions Loughborough University 2009 Human embryonic stem cells Automation CompacT SelecT Scalability Process control Mechanical Engineering not elsewhere classified 2009-02-17 16:49:52 Journal contribution https://repository.lboro.ac.uk/articles/journal_contribution/Automated_scalable_culture_of_human_embryonic_stem_cells_in_feeder_free_conditions/9545834 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.