Mandible Explant Assay for the Analysis of Meckel's Cartilage Development

Ex vivo explant models are a valuable tool for analyzing organ and tissue morphogenesis, providing the opportunity to manipulate and interrogate specific cellular and/or molecular pathways that may not be possible using conventional methods in vivo. The mandible primordia is a remarkably self-organizing structure that has the ability to develop cartilage, bone, teeth, epithelial tissue, and the tongue when grown in culture ex vivo and closely mimics the development of these structures in vivo. Here we describe a robust protocol for the culture of mandibular explants using serum-free, chemically defined culture media. We also describe methods for manipulating mandible and/or Meckel's cartilage development by implantation of agarose beads soaked in various molecular factors to augment mandible development, as well as methods for Alcian blue staining of Meckel's cartilage and immunohistochemistry. This culture method can also be adapted for other molecular analyses, including addition of small-molecule inhibitors and/or growth factors to the culture media, as well as culturing explants from genetically modified mice.