A chimeric vitronectin: IGF-I protein supports feeder-cell-free and serum-free culture of human embryonic stem cells

Stem Cells and Development
Kerry J MantonZee Upton

Abstract

The therapeutic use of human embryonic stem (hES) cells is severely limited by safety concerns regarding their culture in media containing animal-derived or nondefined factors and on animal-derived feeder cells. Thus, there is a pressing need to develop culture techniques that are xeno-free, fully defined, and synthetic. Our laboratory has discovered that insulin-like growth factor (IGF) and vitronectin (VN) bind to each other resulting in synergistic short-term functional effects in several cell types, including keratinocytes and breast epithelial cells. We have further refined this complex into a single chimeric VN:IGF-I protein that functionally mimics the effects obtained upon binding of IGF-I to VN. The aim of the current study was to determine whether hES cells can be serially propagated in feeder-cell-free and serum-free conditions using medium containing our novel chimeric VN:IGF-I protein. Here we demonstrate that hES cells can be serially propagated and retain their undifferentiated state in vitro for up to 35 passages in our feeder-cell-free, serum-free, chemically defined media. We have utilized real-time polymerase chain reaction (PCR), immunofluorescence, and fluorescence-activated cell sorter (FACS) analysis to s...Continue Reading

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Jun 24, 2010·Cell Stem Cell·Kouichi HasegawaMartin F Pera
Jun 3, 2014·Expert Opinion on Biological Therapy·Anastasia G EfthymiouManfred Boehm
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Nov 16, 2010·Stem Cell Research·Andrew B J ProwseTrent P Munro
Jan 1, 2013·Cell Regeneration·Jack W LambsheadAndrew L Laslett

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