Ferguson M W, Honig L S, Slavkin H C
Anat Rec. 1984 Jun;209(2):231-49. doi: 10.1002/ar.1092090210.
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.
从胚胎期的短吻鳄、鸡和小鼠中取出腭突,在发育的不同阶段进行外植,并在化学成分明确的无血清培养基或添加了10%胎牛血清的相同培养基中进行器官培养。每种脊椎动物的腭突单独培养或相互接触培养,还进行了不同动物腭突的异种组合:鸡/小鼠、鸡/短吻鳄和小鼠/短吻鳄。通过活体染色、组织学和扫描电子显微镜对上皮分化(特别是内侧腭突边缘的分化)进行检测。体外内侧边缘上皮分化以及腭闭合机制与每种物种在体内正常观察到的情况相同,即短吻鳄中呈鹅卵石状迁移的上皮、小鼠中的细胞死亡和融合,以及鸡中的角质化和腭裂。在化学成分明确的无血清培养基中分化最佳,且在所有三个物种中均与腭突接触无关。没有异种组合的腭突相互闭合:显然,小鼠和短吻鳄的腭闭合模式差异足够大,以防止它们形成嵌合腭,同时两者都不能诱导共培养的鸡腭突闭合。这些统一的明确培养条件使得大量上皮-间充质重组研究以及特定抑制剂、致畸剂和激素研究成为可能。