Induction of neural crest cells from mouse embryonic stem cells in a serum-free monolayer culture

Yuko Aihara, Yohei Hayashi, Mitsuhi Hirata, Nobutaka Ariki, Shinsuke Shibata, Narihito Nagoshi, Mio Nakanishi, Kiyoshi Ohnuma, Masaki Warashina, Tatsuo Michiue, Hideho Uchiyama, Hideyuki Okano, Makoto Asashima, Miho Kusuda Furue

Research output: Contribution to journalArticlepeer-review

28 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)1287-1294
Number of pages8
JournalInternational Journal of Developmental Biology
Issue number8-9
Publication statusPublished - 2010


  • BMP-4
  • Defined serum-free condition
  • Embryonic stem cell
  • Neural crest

ASJC Scopus subject areas

  • Embryology
  • Developmental Biology


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