RRC ID 37924
著者 Tomizawa M, Shinozaki F, Motoyoshi Y, Sugiyama T, Yamamoto S, Ishige N.
タイトル Involvement of the Wnt signaling pathway in feeder‑free culture of human induced pluripotent stem cells.
ジャーナル Mol Med Rep
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.
巻・号 12(5)
ページ 6797-800
公開日 2015-11-1
DOI 10.3892/mmr.2015.4314
PMID 26398905
MeSH Cells, Cultured Gene Expression Genes, Reporter Humans Induced Pluripotent Stem Cells / physiology* Luciferases / biosynthesis Luciferases / genetics Transcriptional Activation Tubulin / genetics Tubulin / metabolism Wnt Signaling Pathway* beta Catenin / genetics beta Catenin / metabolism
IF 2.1
引用数 1
WOS 分野 MEDICINE, RESEARCH & EXPERIMENTAL ONCOLOGY
リソース情報
ヒト・動物細胞 201B7(HPS0063)