title: Activation of Wnt/ß-catenin signaling in ESC promotes rostral forebrain differentiation in vitro.
abstract: Wnt/ß-catenin signaling is crucial for maintenance of pluripotent state of embryonic stem cell (ESC). However, it is unclear how Wnt/ß-catenin signaling affects the differentiation ability of ESC, especially with regard to rostral forebrain cells. Here, using Rax, rostral forebrain marker, and Wnt/ß-catenin reporter lines, we report ratio of Rax(+) and Wnt responding tissue (Wnt(+)) patterns, which were affected by seeding number of ESC in three-dimensional culture system. Surprisingly, we found ß-catenin level and localization are heterogeneous in ESC colony by immunostaining and time-laps imaging of ß-catenin-mEGFP signals. Moreover, activation of Wnt signaling in ESC promoted expression level and nuclear localization of ß-catenin, and mRNA levels of Wnt antagonists, axin2 and dkk1, leading to upregulating Wnt/ß-catenin reporter in ESC state and Rax expression at differentiation culture day 7. Together, our results suggest that activation of Wnt signaling in ESC promotes the differentiation efficacy of rostral forebrain cells. Wnt-priming culture method may provide a useful tool for applications in the areas of basic science and molecular therapeutics for regenerative medicine.
authors.affiliation: Laboratory for in vitro Histogenesis, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuou-ku, Kobe, Hyogo, 650-0047, Japan. nozomutakata@cdb.riken.jp.
authors.forename: Nozomu
authors.initials: N
authors.lastname: Takata
authors.affiliation: Laboratory for in vitro Histogenesis, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuou-ku, Kobe, Hyogo, 650-0047, Japan.
authors.forename: Eriko
authors.initials: E
authors.lastname: Sakakura
authors.affiliation: Laboratory for Organogenesis and Neurogenesis, RIKEN Center for Developmental Biology, 2-2-3 Minatojima-minamimachi, Chuou-ku, Kobe, Hyogo, 650-0047, Japan.
authors.forename: Yoshiki
authors.initials: Y
authors.lastname: Sasai
mesh_terms: D000818:Animals; D060466:Axin Protein / Q000235:genetics; D002454:Cell Differentiation / Q000502:physiology*; D019070:Cell Lineage / Q000235:genetics; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology*; D004789:Enzyme Activation; D005136:Eye Proteins / Q000378:metabolism; D005434:Flow Cytometry; D018398:Homeodomain Proteins / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics; D051379:Mice; D016548:Prosencephalon / Q000166:cytology*; D012333:RNA, Messenger / Q000235:genetics; D022661:RNA, Untranslated / Q000235:genetics; D014157:Transcription Factors / Q000378:metabolism; D051153:Wnt Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D060449:Wnt Signaling Pathway; D051176:beta Catenin / Q000378:metabolism*
chemical_list: D060466:Axin Protein; C111729:Axin2 protein, mouse; C501127:Dkk1 protein, mouse; D005136:Eye Proteins; C110853:Gt(ROSA)26Sor non-coding RNA, mouse; D018398:Homeodomain Proteins; D036341:Intercellular Signaling Peptides and Proteins; D012333:RNA, Messenger; D022661:RNA, Untranslated; C105441:Rax protein, mouse; D014157:Transcription Factors; D051153:Wnt Proteins; D051176:beta Catenin
keywords: Embryonic stem cells; Rax; Regenerative medicine; Rostral forebrain; Wnt antagonist; Wnt/ß-catenin signaling