Abstract
Neural crest cells are a critical source of many cell types of the vertebrate body. However, as a stem cell population they are peculiar because of the transient nature of their stem cell niche; soon after the multipotent neural crest cells are specified in the neuroepithelium, they become mesenchymal cells that migrate into various destinations in early embryos. These rapid in vivo changes during neural crest development complicate the studies on their stem cell properties. Crestospheres are in vitro maintained primary cultures of premigratory neural crest cells that maintain a mixture of neural crest stem and progenitor cells for weeks without spontaneous differentiation, including the multipotent neural crest stem cells. Here, we describe how crestosphere cultures are initiated from either cranial or trunk levels of chick embryos. Alternatively, the same culture conditions can be used to maintain human embryonic stem cell-derived neural crest cells as crestospheres. Thus, crestospheres provide a useful tool for studies on neural crest stemness.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Simoes-Costa M, Bronner ME (2016) Reprogramming of avian neural crest axial identity and cell fate. Science 352(6293):1570–1573. https://doi.org/10.1126/science.aaf2729
Dupin E, Calloni GW, Coelho-Aguiar JM, Le Douarin NM (2018) The issue of the multipotency of the neural crest cells. Dev Biol. https://doi.org/10.1016/j.ydbio.2018.03.024
Kerosuo L, Nie S, Bajpai R, Bronner ME (2015) Crestospheres: long-term maintenance of multipotent, premigratory neural crest stem cells. Stem Cell Reports 5(4):499–507. https://doi.org/10.1016/j.stemcr.2015.08.017
Cohen AM, Konigsberg IR (1975) A clonal approach to the problem of neural crest determination. Dev Biol 46(2):262–280
Sieber-Blum M, Cohen AM (1980) Clonal analysis of quail neural crest cells: they are pluripotent and differentiate in vitro in the absence of noncrest cells. Dev Biol 80(1):96–106
Lahav R, Ziller C, Dupin E, Le Douarin NM (1996) Endothelin 3 promotes neural crest cell proliferation and mediates a vast increase in melanocyte number in culture. Proc Natl Acad Sci U S A 93(9):3892–3897
Trentin A, Glavieux Pardanaud C, Le Douarin N, Dupin E (2004) Self-renewal capacity is a widespread property of various types of neural crest precursor cells. Proc Natl Acad Sci U S A 101(13):4495–4500
Kerosuo L, Bronner ME (2016) cMyc regulates the size of the premigratory neural crest stem cell pool. Cell Rep 17(10):2648–2659. https://doi.org/10.1016/j.celrep.2016.11.025
Mohlin S, Kunttas E, Persson CU, Abdel-Haq R, Castillo A, Murko C, Bronner ME, Kerusou L (2018) Maintaining trunk neural crest cells as crestospheres. bioRxiv. https://doi.org/10.1101/391599
Reynolds BA, Weiss S (1992) Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 255(5052):1707–1710
Bajpai R, Chen D, Rada Iglesias A, Zhang J, Xiong Y, Helms J, Chang C-P, Zhao Y, Swigut T, Wysocka J (2010) CHD7 cooperates with PBAF to control multipotent neural crest formation. Nature 463(7283):958–962
Martinez-Morales P, Diez Del Corral R, Olivera-Martnez I, Quiroga A, Das R, Barbas J, Storey K, Morales A (2011) FGF and retinoic acid activity gradients control the timing of neural crest cell emigration in the trunk. J Cell Biol 194(3):489–503
Kerosuo L, Bronner M (2014) Biphasic influence of Miz1 on neural crest development by regulating cell survival and apical adhesion complex formation in the developing neural tube. Mol Biol Cell 25(3):347–355
Acloque H, Wilkinson DG, Nieto MA (2008) Chapter 9 in situ hybridization analysis of chick embryos in whole-mount and tissue sections. In: Marianne B-F (ed) Methods in cell biology, vol 87. Academic, New York, pp 169–185. https://doi.org/10.1016/S0091-679X(08)00209-4
Acknowledgments
This work was funded by the Swedish Childhood Cancer Foundation, Thelma Zoéga’s Foundation, Hans von Kantzow’s Foundation, the Mary Béve Foundation, the Royal Physiographic Society of Lund, Magnus Bergvalls Stiftelse (to S.M.) and the Academy of Finland, Sigrid Juselius Foundation, and Ella and Georg Ehrnrooth’s Foundation (to L.K.), and in part by the Division of Intramural Research of the National Institute of Dental and Craniofacial Research at the National Institutes of Health, Department of Health and Human Services.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media New York
About this protocol
Cite this protocol
Mohlin, S., Kerosuo, L. (2018). In Vitro Maintenance of Multipotent Neural Crest Stem Cells as Crestospheres. In: Turksen, K. (eds) Stem Cell Niche. Methods in Molecular Biology, vol 2002. Humana, New York, NY. https://doi.org/10.1007/7651_2018_180
Download citation
DOI: https://doi.org/10.1007/7651_2018_180
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9507-3
Online ISBN: 978-1-4939-9508-0
eBook Packages: Springer Protocols