title: Maintenance of pluripotency in human embryonic stem cells cultured on a synthetic substrate in conditioned medium.
abstract: Realizing the potential clinical and industrial applications of human embryonic stem cells (hESCs) is limited by the need for costly, labile, or undefined growth substrates. Here we demonstrate that trypsin passaging of the hESC lines, HUES7 and NOTT1, on oxygen plasma etched tissue culture polystyrene (PE-TCPS) in conditioned medium is compatible with pluripotency. This synthetic culture surface is stable at room temperature for at least a year and is readily prepared by placing polystyrene substrates in a radio frequency oxygen plasma generator for 5 min. Modification of the polystyrene surface chemistry by plasma etching was confirmed by X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS), which identified elemental and molecular changes as a result of the treatment. Pluripotency of hESCs cultured on PE-TCPS was gauged by consistent proliferation during serial passage, expression of stem cell markers (OCT4, TRA1-60, and SSEA-4), stable karyotype and multi-germlayer differentiation in vitro, including to pharmacologically responsive cardiomyocytes. Generation of cost-effective, easy-to-handle synthetic, defined, stable surfaces for hESC culture will expedite stem cell use in biomedical applications.
authors.affiliation: School of Pharmacy, University of Nottingham, Nottingham, UK.
authors.forename: Magdalena M
authors.initials: MM
authors.lastname: Mahlstedt
authors.forename: David
authors.initials: D
authors.lastname: Anderson
authors.forename: James S
authors.initials: JS
authors.lastname: Sharp
authors.forename: Roger
authors.initials: R
authors.lastname: McGilvray
authors.forename: Maria D Barbadillo
authors.initials: MD
authors.lastname: Muñoz
authors.forename: Lee D
authors.initials: LD
authors.lastname: Buttery
authors.forename: Morgan R
authors.initials: MR
authors.lastname: Alexander
authors.forename: Felicity R A J
authors.initials: FR
authors.lastname: Rose
authors.forename: Chris
authors.initials: C
authors.lastname: Denning
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D017077:Culture Media, Conditioned / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D039904:Pluripotent Stem Cells / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D013499:Surface Properties
chemical_list: D017077:Culture Media, Conditioned