Development of a suspension organ culture of the fetal rat palate.

On the basis of an already established suspension organ culture system of mouse palate anlagen, we developed a corresponding culture system for rat palate anlagen. In order to optimize the culture results we systematically studied the influence of main "culture conditions" such as dissection technique, rotation speed, gassing schedule, and developmental stage at the onset of culture for mice and rat palate anlagen. This system allows culturing rat palate anlagen from day 15 of gestation to day 18 + 8 h (80 h) under serum- and antibiotic-free conditions using a chemically defined medium, resulting in 90% fused palates. The explants, containing the maxillary vault and the palatal shelves, were cultured in siliconized culture flasks at a rotation speed of 12 rpm and a temperature of 37 degrees C (Table 1).