Dissociation of frog olfactory epithelium

We report a method for producing cell suspensions from frog olfactory epithelium. The tissue is incubated for 45 min at room temperature in a solution which causes dissociation of the epithelium. The solution is an isotonic saline buffered to maintain a pH of 10.3 and a free Ca2+ ion concentration of 10−6 M. The method uses no degradative enzymes, except for a brief DNase treatment. The resulting cell suspension contains single olfactory receptor neurons, sustentacular cells, glandular cells, and respiratory epithelial cells. The cells are viable as judged by vital staining, ciliary motility, and synthesis of RNA. Some types of cells lose their normal columnar shapes and become rounded in the suspension.