Protocol | Published:

Primary culture of chick, mouse or human neural crest cells

Nature Protocols volume 6, pages 15681577 (2011) | Download Citation

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Abstract

A highly enriched population of neural crest cells (NCCs) from amniote embryos, such as from chicks, mice and humans, is desirable for experiments in fate determination. NCCs are also useful for testing the functional effects of molecular changes underlying numerous human diseases of neural crest derivatives and for investigating their potential for therapeutic compensation. This protocol details embryonic microdissection followed by neural tube explantation. Conditions favoring NCC expansion and the maintenance of their stem cell–like properties are described. Although neural crest–like cells can be derived from a number of sites in the mature organism, full potential is best ensured by their purification from their source tissue at the outset of migration. Going from embryo to established cell line takes 4 d; the first is the most labor-intensive day, but minimal intervention is required thereafter.

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Change history

  • 11 October 2013

     In the version of this article initially published, the descriptions of how to make FGF and complete NCC culture medium were incorrect. The text initially read "FGF (2.5 µg ml–1): Use 4 ml 10% (wt/vol) BSA solution to dissolve 25 µg of FGF." In addition, in the details for making complete NCC culture medium, the text initially read "Add 1 µl of EGF (2.5 µg ml–1) and 8 µl of FGF (10 µg ml–1)." The phrase about FGF should have read "FGF (2.5 µg ml–1): Use 4 ml 10% (wt/vol) BSA solution to dissolve 10 µg of FGF." For complete NCC culture medium, the text should have read "Add 1 µl of EGF (10 µg ml–1) and 8 µl of FGF (2.5 µg ml–1)." The errors have been corrected in the HTML and PDF versions of the article.

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Acknowledgements

H.E. has been supported by the Sturge-Weber foundation (2001), the INSERM Avenir program (2002–2005), the Fondation pour la Recherche Médicale (DEQ20071210511), the Agence Nationale pour la Recherche (ANR2007-CRANIRARE) and Nevus Outreach. In addition to all the authors who have developed the neural crest culture protocols referenced herein and to the directors of INSERM U781 and U910 who have made it possible for me to obtain embryonic tissues over the years, I extend my particular gratitude to E. Dupin, C. Glavieux-Pardanaud, A. Gonçalves-Trentin, S. Thomas and C. Ziller for aiding me in optimizing conditions.

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Affiliations

  1. Institut National de la Santé et de la Recherche Médicale (INSERM) U910, Université de la Méditerranée Faculté de Médecine, Marseille, France.

    • Heather Etchevers

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The author declares no competing financial interests.

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Correspondence to Heather Etchevers.

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DOI

https://doi.org/10.1038/nprot.2011.398

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