Development of the Fetal Mouse Palate in Suspension Organ Culture

Abstract

Explanted palates of day 12 and day 13 mouse fetuses were cultured in a chemically defined serumless medium for 48–72 h by a suspension culture technique. The palate of day 12 fetuses closed successfully within 72 h and that of day 13 fetuses within 48 h. Both macroscopically and histologically the in vitro fusion of palatal shelves simulated the palatogenetic process in vivo. This novel technique for culturing the fetal mouse palate may be of potential use for the study of palatogenesis and in developmental toxicology.

Programmed cell death is not a necessary prerequisite for fusion of the fetal mouse palate.

“…1A). Thus, the process of macroscopic palatal fusion in vitro basically simulated that occurring in vivo, as observed previously (Shiota et al, 1990). Grossly, palatal fusion was observed in 92% of the palates in the control group cultured for 24 hours.…”

“…We confirmed that the suspension culture of fetal mouse palates in roller bottles (Shiota et al, 1990) was superior to static culture systems where explants were placed on Millipore filters and subjected to submerged culture. In such static cultures, nonphysiological massive cell death occurred in explants and the cultured tissues easily became necrotic (data not shown).…”

“…In such static cultures, nonphysiological massive cell death occurred in explants and the cultured tissues easily became necrotic (data not shown). In contrast, such non-specific cell death and necrosis were minimal in suspension organ culture (Shiota et al, 1990). Thus, the suspension organ culture of fetal mouse palates and the use of caspase inhibitors and ATA have enabled to prevent the occurrence of cell death during palatal fusion and to examine its roles in murine palatogenesis.…”

The regulation of endogenous retinoic acid level through CYP26B1 is required for elevation of palatal shelves

“…To understand whether the intrinsic factors are impaired in Cyp26b1À/À fetuses, we performed fetal palate culture in suspension (Shiota et al, 1990) (Fig. 5A-D).…”

“…Fetal Mouse Palate in Suspension Organ Culture E13.5 palatal shelves were cultivated in suspension for 20 hr as previously described (Shiota et al, 1990). Briefly, they were placed into a sterile 15-ml glass bottle containing 1.5 ml of BGJ-b (Gibco-Life Technologies, Gaithersburg, MD), 6 mg/ml of BSA, and 150 mg/ml of ascorbic acid.…”