Heparin promotes the growth of human embryonic stem cells in a defined serum-free medium.
A major limitation in developing applications for the use of human embryonic stem cells (HESCs) is our lack of knowledge of their responses to specific cues that control self-renewal, differentiation, and lineage selection. HESCs are most commonly maintained on inactivated mouse embryonic fibroblast feeders in medium supplemented with FCS, or proprietary replacements such as knockout serum-replacement together with FGF-2. These undefined culture conditions hamper analysis of the mechanisms that control HESC behavior. We have now developed a defined serum-free medium, hESF9, for the culture of HESCs on a type I-collagen substrate without feeders. In contrast to other reported media for the culture of HESCs, this medium has a lower osmolarity (292 mosmol/liter), l-ascorbic acid-2-phosphate (0.1 microg/ml), and heparin. Insulin, transferrin, albumin conjugated with oleic acid, and FGF-2 (10 ng/ml) were the only protein components. Further, we found that HESCs would proliferate in the absence of exogenous FGF-2 if heparin was also present. However, their growth was enhanced by the addition of FGF-2 up to 10 ng/ml although higher concentrations were deleterious in the presence of heparin.
Furue
Miho K
MK
JCRB Cell Bank, Division of Bioresources, National Institute of Biomedical Innovation, Osaka 567-0085, Japan.
Na
Jie
J
Jackson
Jamie P
JP
Okamoto
Tetsuji
T
Jones
Mark
M
Baker
Duncan
D
Hata
Ryu-Ichiro
R
Moore
Harry D
HD
Sato
J Denry
JD
Andrews
Peter W
PW
D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D049109:Cell Proliferation; D016895:Culture Media, Serum-Free; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects*; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D006493:Heparin / Q000494:pharmacology*; D006801:Humans; D015398:Signal Transduction
D016895:Culture Media, Serum-Free; D005346:Fibroblast Growth Factors; D006493:Heparin