title: Automated, scalable culture of human embryonic stem cells in feeder-free conditions.
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.
authors.affiliation: Loughborough University, UK. r.j.thomas@lboro.ac.uk
authors.forename: Rob J
authors.initials: RJ
authors.lastname: Thomas
authors.forename: David
authors.initials: D
authors.lastname: Anderson
authors.forename: Amit
authors.initials: A
authors.lastname: Chandra
authors.forename: Nigel M
authors.initials: NM
authors.lastname: Smith
authors.forename: Lorraine E
authors.initials: LE
authors.lastname: Young
authors.forename: David
authors.initials: D
authors.lastname: Williams
authors.forename: Chris
authors.initials: C
authors.lastname: Denning
mesh_terms: D015415:Biomarkers / Q000032:analysis; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D053595:Embryonic Stem Cells; D004867:Equipment Design; D006801:Humans; D007621:Karyotyping; D008856:Microscopy, Fluorescence; D039904:Pluripotent Stem Cells; D012371:Robotics; D014357:Trypsin / Q000737:chemistry
chemical_list: D015415:Biomarkers; D014357:Trypsin