Open AccessSecreted Proteoglycans Directly Mediate Human Embryonic Stem Cell‐Basic Fibroblast Growth Factor 2 Interactions Critical for Proliferation
Author(s) -
Levenstein Mark E.,
Berggren W. Travis,
Lee Ji Eun,
Conard Kevin R.,
Llanas Rachel A.,
Wagner Ryan J.,
Smith Lloyd M.,
Thomson James A.
Publication year - 2008
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1634/stemcells.2007-1056
Subject(s) - embryonic stem cell , biology , fibroblast growth factor , microbiology and biotechnology , fibroblast , cell culture , cell growth , stem cell , cell , growth factor , biochemistry , genetics , gene , receptor
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.