Articles Cell Growth/Differentiation/Apoptosis

In Vitro Cellular & Developmental Biology - Animal

, Volume 41, Issue 1, pp 19-28

Leukemia inhibitory factor as an anti-apoptotic mitogen for pluripotent mouse embryonic stem cells in a serum-free medium without feeder cells

  • Miho FurueAffiliated withDepartment of Biochemistry and Molecular Biology, Kanagawa Dental College Email author 
  • , Tetsuji OkamotoAffiliated withDepartment of Molecular Oral Medicine and Maxillofacial Surgery, Division of Frontier Medical Science, Graduate School of Biomedical Sciences, Hiroshima University
  • , Yohei HayashiAffiliated withDepartment of Life Sciences (Biology), Graduate School of Arts and Sciences, The University of Tokyo
  • , Hitoshi OkochiAffiliated withDepartment of Tissue Regeneration, Research Institute, International Medical Center of Japan
  • , Manabu FujimotoAffiliated withDepartment of Tissue Regeneration, Research Institute, International Medical Center of Japan
  • , Yasufumi MyoishiAffiliated withDepartment of Molecular Oral Medicine and Maxillofacial Surgery, Division of Frontier Medical Science, Graduate School of Biomedical Sciences, Hiroshima University
  • , Takanori AbeAffiliated withDepartment of Biological Science, Graduate School of Science, The University of Tokyo
  • , Kiyoshi OhnumaAffiliated withDepartment of Life Sciences (Biology), Graduate School of Arts and Sciences, The University of Tokyo
  • , Gordon H. SatoAffiliated withMinistry of Fisheries
    • , Makoto AsashimaAffiliated withDepartment of Life Sciences (Biology), Graduate School of Arts and Sciences, The University of TokyoDepartment of Biological Science, Graduate School of Science, The University of TokyoInternational Cooperative Research Project (ICORP)-Japan Science and Technology Agency (JST), The University of Tokyo
    • , J. Denry SatoAffiliated withMarine Cell Line and Stem Cell Program, Mount Desert Island Biological Laboratory

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Summary

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 MBP4 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.

Key words

ES mouse embryonic stem cells serum-free LIF nanog Oct-3/4