小鼠味蕾细胞分离及体外培养方法

The Isolation and in Vitro Culture of Mouse Taste Bud Cells

以ICR小鼠的味觉上皮为材料,优化味觉上皮及味蕾细胞分离方法,将分离的味细胞置于Ⅰ型鼠尾胶原包被的含有IMDM培养基的培养板上培养,相差显微镜下观察味蕾细胞形态变化。免疫组织化染色和荧光染色用于鉴定味蕾细胞。结果显示味蕾细胞离体培养一段时间后形态异于体内,但仍具味蕾细胞特有的与细胞骨架和细胞内信号相关的分子标记如:α-gustducin、细胞角蛋白8。由此证明这是一种有效的小鼠的味蕾细胞分离及体外培养方法,为离体研究味蕾细胞生理生化特性提供了更好的途径。

The lingual epithelium of ICR mouse was utilized for this study. The isolated taste bud cells were cultured in rat tail collagen type Ⅰ-coated cultured plate with IMDM. Phase contrast microscopy was used to record their morphological changes during culture. Immunohistochemistry and immunofluorescent staining was performed to identify taste bud cells. Results showed that taste bud cells changed their morphology after culturing for more than one week, but still retained such molecular markers as： 1） α-gustducin which is involved in intracellular signaling, 2） cytokeratin 8 which is a component of cytoskeleton. These results demonstrated that a primary method might be thus established to separate taste bud cells from the circumvallate of ICR mouse and to culture them in vitro so as to provide an effective way to study their physiological and biochemical characteristics.