In Vitro Maintenance of Multipotent Neural Crest Stem Cells as Crestospheres

  • Sofie Mohlin
  • Laura KerosuoEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2002)


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.


Chick embryo Cranial neural crest Crestospheres Human ES cell-derived neural crest Multipotency Neural crest Self-renewal Trunk neural crest 



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.


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Copyright information

© Springer Science+Business Media New York 2018

Authors and Affiliations

  1. 1.Division of Pediatrics, Department of Clinical SciencesLund UniversityLundSweden
  2. 2.Department of Health and Human ServicesNeural Crest Development and Disease Unit, National Institute of Dental and Craniofacial Research, National Institutes of HealthBethesdaUSA
  3. 3.Biochemistry and Developmental Biology, Faculty of MedicineUniversity of HelsinkiHelsinkiFinland

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