Differentiation of cultured palatal shelves from alligator, chick, and mouse embryos

Palatal shelves from embryonic alligators, chicks, and mice were explanted at various stages of development and organ cultured in either chemically defined, serumless media or the same media supplemented with 10% fetal calf serum. Shelves from each vertebrate were either cultured singly or in contact, and heterologous combinations of palatal shelves from different animals were made: chick/mouse, chick/alligator, and mouse/alligator. Epithelial differentiation (particularly that of the medial shelf edge) was assayed by vital staining, histology, and scanning electron microscopy. In vitro medial edge epithelial differentiation, and consequently the mechanisms of palatal closure, were identical to those normally seen in vivo for each species, i.e., cobble-stoned migrating epithelia in the alligator, cell death and fusion in the mouse, and keratinisation and cleft palate in the chick. Differentiation was optimal in the chemically defined, serumless media and was independent of shelf contact in all three species. No heterologous combinations of palatal shelves closed with each other: Evidently, the modes of palatal closure in mice and alligators are sufficiently different to prevent them forming a chimeric palate, whilst neither is capable of inducing closure in a cocultured chick palatal shelf. These unified defined culture conditions make possible a large number of epithelial-mesenchymal recombination studies as well as specific inhibitor, teratogenic, and hormonal investigations.