Dissection of Xenopus laevis neural crest for in vitro explant culture or in vivo transplantation

• Published in: Journal of visualized experiments 2014-03 (85)
• Main Authors: Milet, Cecile, Monsoro-Burq, Anne Helene
• Format: Article
• Language: English
• Subjects: Animals; Developmental Biology; Dissection; Dissection - methods; Epithelial-Mesenchymal Transition; Life Sciences; Neural Crest; Neural Crest - cytology; Neural Crest - surgery; Neural Crest - transplantation; Organ Culture Techniques; Organ Culture Techniques - methods; Xenopus laevis; Xenopus laevis - embryology; Xenopus laevis - surgery
• ISSN: 1940-087X; 1940-087X
• DOI: 10.3791/51118

The neural crest (NC) is a transient dorsal neural tube cell population that undergoes an epithelium-to-mesenchyme transition (EMT) at the end of neurulation, migrates extensively towards various organs, and differentiates into many types of derivatives (neurons, glia, cartilage and bone, pigmented and endocrine cells). In this protocol, we describe how to dissect the premigratory cranial NC from Xenopus laevis embryos, in order to study NC development in vivo and in vitro. The frog model offers many advantages to study early development; abundant batches are available, embryos develop rapidly, in vivo gain and loss of function strategies allow manipulation of gene expression prior to NC dissection in donor and/or host embryos. The NC explants can be plated on fibronectin and used for in vitro studies. They can be cultured for several days in a serum-free defined medium. We also describe how to graft NC explants back into host embryos for studying NC migration and differentiation in vivo.