We have developed a serum-free medium, designated ESF7, in which leukemia inhibitory factor (LIF) clearly stimulated murine embryonic stem (ES) cell proliferation accompanied by increased expression of nanog and Rex-1 and decreased FGF-5 expression. These effects were dependent on the concentration of LIF. The ES cells maintained in ESF7 medium for more than 2 yr retained an undifferentiated phenotype, as manifested by the expression of the transcription factor Oct-3/4, the stem cell marker SSEA-1, and alkaline phosphatase. Withdrawal of LIF from ESF7 medium resulted in ES cell apoptosis. Addition of serum to ESF7 medium promoted ES cell differentiation. Addition of BMP4 promoted ES cell differentiation into simple epithelial-like cells. In contrast, FGF-2 promoted ES cell differentiation into neuronal and glial-like cells. Under serum-free culture conditions, LIF was sufficient to stimulate cell proliferation, it inhibited cell differentiation, and it maintained self-renewal of ES cells. Because this simple serum-free adherent monoculture system supports the long-term propagation of pluripotent ES cells in vitro, it will allow the elucidation of ES cell responses to growth factors under defined conditions.
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1 January 2005
LEUKEMIA INHIBITORY FACTOR AS AN ANTI-APOPTOTIC MITOGEN FOR PLURIPOTENT MOUSE EMBRYONIC STEM CELLS IN A SERUM-FREE MEDIUM WITHOUT FEEDER CELLS
MIHO FURUE,
TETSUJI OKAMOTO,
YOHEI HAYASHI,
HITOSHI OKOCHI,
MANABU FUJIMOTO,
YASUFUMI MYOISHI,
TAKANORI ABE,
KIYOSHI OHNUMA,
GORDON H. SATO,
MAKOTO ASASHIMA,
J. DENRY SATO
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In Vitro Cellular & Developmental Biology - Animal
Vol. 41 • No. 1
January 2005
Vol. 41 • No. 1
January 2005
ES
LIF
mouse embryonic stem cells
nanog
Oct-3/4
serum-free