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Article

Involvement of the Wnt signaling pathway in feeder‑free culture of human induced pluripotent stem cells

  • Authors:
    • Minoru Tomizawa
    • Fuminobu Shinozaki
    • Yasufumi Motoyoshi
    • Takao Sugiyama
    • Shigenori Yamamoto
    • Naoki Ishige
  • View Affiliations / Copyright

    Affiliations: Department of Gastroenterology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 284‑0003, Japan, Department of Radiology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 284‑0003, Japan, Department of Neurology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 284‑0003, Japan, Department of Rheumatology, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 284‑0003, Japan, Department of Pediatrics, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 284‑0003, Japan, Department of Neurosurgery, National Hospital Organization, Shimoshizu Hospital, Yotsukaido, Chiba 284‑0003, Japan
  • Pages: 6797-6800
    |
    Published online on: September 10, 2015
       https://doi.org/10.3892/mmr.2015.4314
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Abstract

Activin A maintains the pluripotency of human induced pluripotent stem (hiPS) cells. A combination of activin A and CHIR99021 (CHIR), a specific inhibitor of glycogen synthase‑3β, is suitable for feeder‑free culture of hiPS cells. In the present study, the specific role of the Wnt signaling pathway in cells cultured under different conditions was investigated. Following transfection with the reporter plasmids, TOPflash and FOPflash, hiPS cells were cultured in medium, containing activin A, CHIR, leukemia inhibitory factor (LIF) or SB431542, a specific inhibitor of activin A. A luciferase reporter assay was performed 48 h later. Western blot analysis was performed to determine the expression levels of β‑catenin and tubulin‑α. The activity of Wnt in hiPS cells was suppressed by culture in the presence of activin A. The activation of the Wnt pathway was most marked when the cells were cultured with a combination of activin A and CHIR. Addition of SB431542 into the culture revealed no significant change in the Wnt pathway. Western blot analysis revealed that β‑catenin accumulated most often in cells cultured with activin A and CHIR. β‑catenin also accumulated in cells cultured with activin A alone. Culture with activin A and CHIR most effectively stimulated the Wnt signaling pathway, as measured by luciferase assays using TOPflash and FOP flash as reporter plasmids. β‑catenin accumulated in the hiPS cells cultured with activin A, via a mechanism, which remains to be elucidated. The Wnt signaling pathway may be important for hiPS cell growth in feeder‑free culture.
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Copy and paste a formatted citation
Spandidos Publications style
Tomizawa M, Shinozaki F, Motoyoshi Y, Sugiyama T, Yamamoto S and Ishige N: Involvement of the Wnt signaling pathway in feeder‑free culture of human induced pluripotent stem cells. Mol Med Rep 12: 6797-6800, 2015.
APA
Tomizawa, M., Shinozaki, F., Motoyoshi, Y., Sugiyama, T., Yamamoto, S., & Ishige, N. (2015). Involvement of the Wnt signaling pathway in feeder‑free culture of human induced pluripotent stem cells. Molecular Medicine Reports, 12, 6797-6800. https://doi.org/10.3892/mmr.2015.4314
MLA
Tomizawa, M., Shinozaki, F., Motoyoshi, Y., Sugiyama, T., Yamamoto, S., Ishige, N."Involvement of the Wnt signaling pathway in feeder‑free culture of human induced pluripotent stem cells". Molecular Medicine Reports 12.5 (2015): 6797-6800.
Chicago
Tomizawa, M., Shinozaki, F., Motoyoshi, Y., Sugiyama, T., Yamamoto, S., Ishige, N."Involvement of the Wnt signaling pathway in feeder‑free culture of human induced pluripotent stem cells". Molecular Medicine Reports 12, no. 5 (2015): 6797-6800. https://doi.org/10.3892/mmr.2015.4314
Copy and paste a formatted citation
x
Spandidos Publications style
Tomizawa M, Shinozaki F, Motoyoshi Y, Sugiyama T, Yamamoto S and Ishige N: Involvement of the Wnt signaling pathway in feeder‑free culture of human induced pluripotent stem cells. Mol Med Rep 12: 6797-6800, 2015.
APA
Tomizawa, M., Shinozaki, F., Motoyoshi, Y., Sugiyama, T., Yamamoto, S., & Ishige, N. (2015). Involvement of the Wnt signaling pathway in feeder‑free culture of human induced pluripotent stem cells. Molecular Medicine Reports, 12, 6797-6800. https://doi.org/10.3892/mmr.2015.4314
MLA
Tomizawa, M., Shinozaki, F., Motoyoshi, Y., Sugiyama, T., Yamamoto, S., Ishige, N."Involvement of the Wnt signaling pathway in feeder‑free culture of human induced pluripotent stem cells". Molecular Medicine Reports 12.5 (2015): 6797-6800.
Chicago
Tomizawa, M., Shinozaki, F., Motoyoshi, Y., Sugiyama, T., Yamamoto, S., Ishige, N."Involvement of the Wnt signaling pathway in feeder‑free culture of human induced pluripotent stem cells". Molecular Medicine Reports 12, no. 5 (2015): 6797-6800. https://doi.org/10.3892/mmr.2015.4314
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