Secreted Proteoglycans Directly Mediate Human Embryonic Stem Cell-Basic Fibroblast Growth Factor 2 Interactions Critical for Proliferation
Open Access
- 18 September 2008
- journal article
- research article
- Published by Oxford University Press (OUP) in The International Journal of Cell Cloning
- Vol. 26 (12) , 3099-3107
- https://doi.org/10.1634/stemcells.2007-1056
Abstract
Human embryonic stem (ES) cells can be maintained in an undifferentiated state if the culture medium is first conditioned on a layer of mouse embryonic fibroblast (MEF) feeder cells. Here we show that human ES cell proliferation is coordinated by MEF-secreted heparan sulfate proteoglycans (HSPG) in conditioned medium (CM). These HSPG and other heparinoids can stabilize basic fibroblast growth factor (FGF2) in unconditioned medium at levels comparable to those observed in CM. They also directly mediate binding of FGF2 to the human ES cell surface, and their removal from CM impairs proliferation. Finally, we have developed a purification scheme for MEF-secreted HSPG in CM. Using column chromatography, immunoblotting, and mass spectrometry-based proteomic analysis, we have identified multiple HSPG species in CM. The results demonstrate that HSPG are key signaling cofactors in CM-based human ES cell culture. Disclosure of potential conflicts of interest is found at the end of this article.Keywords
This publication has 34 references indexed in Scilit:
- IGF and FGF cooperatively establish the regulatory stem cell niche of pluripotent human cells in vitroNature, 2007
- Heparan sulfate regulation of progenitor cell fateJournal of Cellular Biochemistry, 2006
- Interactions between heparan sulfate and proteins: the concept of specificityThe Journal of cell biology, 2006
- Specific Capture of Mammalian Cells by Cell Surface Receptor Binding to Ligand Immobilized on Gold Thin FilmsJournal of Proteome Research, 2006
- Insights into the role of heparan sulphate in fibroblast growth factor signallingBiochemical Society Transactions, 2006
- Derivation of human embryonic stem cells in defined conditionsNature Biotechnology, 2006
- Basic Fibroblast Growth Factor Support of Human Embryonic Stem Cell Self-RenewalThe International Journal of Cell Cloning, 2005
- Basic Fibroblast Growth Factor Supports Undifferentiated Human Embryonic Stem Cell Growth Without Conditioned MediumThe International Journal of Cell Cloning, 2005
- Embryonic Stem Cell Lines Derived from Human BlastocystsScience, 1998
- Developmental changes in heparan sulfate expression: in situ detection with mAbs.The Journal of cell biology, 1992