A reliable method to obtain cells of taste buds from fungiform papillae of mice.
Taste buds consist of four kinds of cells which have distinct characteristics and play different roles in recognizing chemical compounds contained in foodstuffs. In this study we describe a procedure for separating viable taste bud cells from the fungiform papillae in mice. After sacrifice with CO(2), the mouse tongue was excised and immediately incubated in collagenase II and dispase II. The epithelium with fungiform papillae was then peeled away from underlying tissue and the anterior one-third region was incubated in a solution of 0.25% trypsin and 0.02M ethylene-diamine-tetraacetic acid (EDTA) for 8-12min. Following incubation, a cell suspension was obtained by mechanical dissociation. Cells in suspension were identified as taste bud cells by their morphology and by immunofluorescence. A 0.25% trypan blue staining demonstrated that nearly 90% of these cells remained viable. Micrographs from scanning electron microscopy illustrated that taste buds were dissociated from the fungiform papillae, while maintaining the integrity of the other part of the dissociated lingual epithelium during incubation. Such a method allows acquisition of viable taste cells and will aid further research in the study of gustatory characteristics.
Qin
Yu-Mei
YM
Zhejiang Gongshang University, Hangzhou, China.
Shi
Jin-Qin
JQ
Zhang
Gen-Hua
GH
Deng
Shao-Ping
SP
Wang
Teng-Hao
TH
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D004848:Epithelium / Q000648:ultrastructure*; D005455:Fluorescent Antibody Technique; D053533:Keratin-8 / Q000378:metabolism; D008297:Male; D051379:Mice; D008813:Mice, Inbred ICR; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D013650:Taste Buds / Q000166:cytology* / Q000378:metabolism
D053533:Keratin-8