title: Induction of neural crest cells from mouse embryonic stem cells in a serum-free monolayer culture.
abstract: The neural crest (NC) is a group of cells located in the neural folds at the boundary between the neural and epidermal ectoderm. NC cells differentiate into a vast range of cells,including neural cells, smooth muscle cells, bone and cartilage cells of the maxillofacial region, and odontoblasts. The molecular mechanisms underlying NC induction during early development remain poorly understood. We previously established a defined serum-free culture condition for mouse embryonic stem (mES) cells without feeders. Here, using this defined condition, we have developed a protocol to promote mES cell differentiation into NC cells in an adherent monolayer culture. We found that adding bone morphogenetic protein (BMP)-4 together with fibroblast growth factor (FGF)-2 shifts mES cell differentiation into the NC lineage. Furthermore, we have established a cell line designated as P0-6 that is derived from the blastocysts of P0-Cre/Floxed-EGFP mice expressing EGFP in an NC-lineage-specific manner. P0-6 cells cultured using this protocol expressed EGFP. This protocol could be used to help clarify the mechanisms by which cells differentiate into the NC lineage and to assist the development of applications for clinical therapy.
authors: Department of Life Sciences (Biology), Graduate School of Arts and Sciences, University of Tokyo, Tokyo, Japan.
authors: Yuko
authors: Y
authors: Aihara
authors: Yohei
authors: Y
authors: Hayashi
authors: Mitsuhi
authors: M
authors: Hirata
authors: Nobutaka
authors: N
authors: Ariki
authors: Shinsuke
authors: S
authors: Shibata
authors: Narihito
authors: N
authors: Nagoshi
authors: Mio
authors: M
authors: Nakanishi
authors: Kiyoshi
authors: K
authors: Ohnuma
authors: Masaki
authors: M
authors: Warashina
authors: Tatsuo
authors: T
authors: Michiue
authors: Hideho
authors: H
authors: Uchiyama
authors: Hideyuki
authors: H
authors: Okano
authors: Makoto
authors: M
authors: Asashima
authors: Miho Kusuda
authors: MK
authors: Furue
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001755:Blastocyst / Q000166:cytology / Q000378:metabolism; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D019070:Cell Lineage / Q000187:drug effects / Q000502:physiology*; D002470:Cell Survival / Q000187:drug effects; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D015870:Gene Expression / Q000187:drug effects; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007381:Intermediate Filament Proteins / Q000235:genetics; D051379:Mice; D008822:Mice, Transgenic; D008856:Microscopy, Fluorescence; D009419:Nerve Tissue Proteins / Q000235:genetics; D064231:Nestin; D009432:Neural Crest / Q000166:cytology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000071250:Snail Family Transcription Factors; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics
chemical_list: D055415:Bone Morphogenetic Protein 4; D016895:Culture Media, Serum-Free; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; D064231:Nestin; D000071250:Snail Family Transcription Factors; D014157:Transcription Factors; D016222:Fibroblast Growth Factor 2; D049452:Green Fluorescent Proteins