Journal
BIOTECHNOLOGY AND BIOENGINEERING
Volume 105, Issue 1, Pages 130-140Publisher
WILEY-BLACKWELL
DOI: 10.1002/bit.22520
Keywords
human embryonic stem cells (hESCs); radio frequency plasma etching; synthetic culture surface; pluripotency; oxygen; nitrogen; XPS; ToF-SIMS
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Funding
- University of Nottingham
- British Heart Foundation
- Medical Research Council
- Biotechnology & Biological Sciences Research Council
- BBSRC [BB/E006159/1] Funding Source: UKRI
- MRC [G0801098, G113/30] Funding Source: UKRI
- Biotechnology and Biological Sciences Research Council [BB/E006159/1] Funding Source: researchfish
- British Heart Foundation [PG/09/027/27141] Funding Source: researchfish
- Medical Research Council [G0801098, G113/30] Funding Source: researchfish
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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. Biotechnol. Bioeng. 2010;105: 130-140. (C) 2009 Wiley Periodicals, Inc.
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