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pp 1-15 | Cite as

Direct Conversion of Mouse Embryonic Fibroblasts into Neural Crest Cells

  • Tsutomu Motohashi
  • Takahiro Kunisada
Protocol
Part of the Methods in Molecular Biology book series

Abstract

Neural crest cells (NCCs) are multipotent cells that emerge from the edges of the neural folds and extensively migrate throughout developing embryos. Dorsolaterally migrating NCCs colonize skin, differentiate into skin melanocytes, and lose their multipotency. Multipotent NCCs or NCCs derived multipotent stem cells (MSCs) were recently detected in their migrated locations, including skin, despite restrictions in cell fate acquisition following migration. Since many features of NCCs have yet to be revealed, the novel properties of NCCs represent an important and interesting field in stem cell biology. We previously reported the direct conversion of mouse embryonic fibroblasts (MEFs) into NCCs by the forced expression of the transcription factors C-MYC, KLF4, and SOX10. We herein describe the methods employed for direct conversion: retrovirus infection for the forced expression of transcription factors, a flow cytometry-sorting method for the isolation of converted NCCs, and culture methods for the maintenance and differentiation of the converted NCCs.

Keywords

C-MYC Direct conversion Flow cytometer KLF-4 Neural crest cells P75 SOX10 

Notes

Acknowledgements

This study was supported by the Gifu University Graduate School of Medicine Research Grant Program, by a research grant from the Japan Science and Technology Agency CREST, and by a grant from the program Grants-in-Aid for Scientific Research (C) from the Japan Society for Promotion for Science. A part of the work was also supported by the Intramural Research Program of the National Institutes of Health, National Institute on Aging, USA.

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Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  1. 1.Department of Tissue and Organ Development, Regeneration and Advanced Medical ScienceGifu University Graduate School of MedicineGifuJapan

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