A novel method to isolate odontoblasts from rat incisor

Historically, odontoblasts have been isolated from rat incisor using a surg ical curette to separate these cells from the dentin. Isolation of odontobl asts using this approach typically resulted in cells with membrane properti es that made the application of patch-clamp electrophysiological techniques prohibitive. The studies here describe a new procedure for isolating matur e odontoblasts from adult rat incisor to obtain enriched populations of int act, viable odontoblasts that can be readily studied using patch-clamp meth odologies. Identification of isolated cells as odontoblasts was confirmed u sing in situ mRNA hybridization for expression of dentin sialoprotein, oste ocalcin, bone sialoprotein, and type I collagen, and calcium flux was monit ored in these cells by means of fura-2 microfluorometry. We suggest that ei ther single odontoblasts or clusters of these cells isolated by this new me thod would be an ideal preparation for the study of odontoblast properties using electrophysiological techniques, in situ hybridization and/or microfl uorometry.