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Proc Natl Acad Sci U S A. 2011 Nov 15;108(46):18714-9. doi: 10.1073/pnas.1114854108. Epub 2011 Nov 7.

Surface-engineered substrates for improved human pluripotent stem cell culture under fully defined conditions.

Author information

1
The Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA.

Abstract

The current gold standard for the culture of human pluripotent stem cells requires the use of a feeder layer of cells. Here, we develop a spatially defined culture system based on UV/ozone radiation modification of typical cell culture plastics to define a favorable surface environment for human pluripotent stem cell culture. Chemical and geometrical optimization of the surfaces enables control of early cell aggregation from fully dissociated cells, as predicted from a numerical model of cell migration, and results in significant increases in cell growth of undifferentiated cells. These chemically defined xeno-free substrates generate more than three times the number of cells than feeder-containing substrates per surface area. Further, reprogramming and typical gene-targeting protocols can be readily performed on these engineered surfaces. These substrates provide an attractive cell culture platform for the production of clinically relevant factor-free reprogrammed cells from patient tissue samples and facilitate the definition of standardized scale-up friendly methods for disease modeling and cell therapeutic applications.

PMID:
22065768
PMCID:
PMC3219112
DOI:
10.1073/pnas.1114854108
[Indexed for MEDLINE]
Free PMC Article

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