In Vitro Maintenance of Multipotent Neural Crest Stem Cells as Crestospheres
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.
KeywordsChick 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.
- 9.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