An odontogenic epithelial cell line, ROE‐2B, was established by propagating disaggregated immature, unmineralized maxillary third molar tooth germs from 11‐day old Sprague‐Dawley rats on a feeder layer of Mitomycin‐C treated NIH 3T3 embryonic mouse fibroblasts. The cell line has been maintained for more than 6 months and through 7 passages. Light microscopic examination of cells revealed colonies with epithelial morphology. Electron microscopic examination confirmed the epithelial nature by the demonstration of tonofilaments, desmosomes and basal lamina. Cells were also shown to have secretory vacuoles, an abundance of granular endoplasmic reticulum, free ribosomes, Golgi complex and mitochondria. Surface activity in the form of pseudopodia‐like projections and micropinocytosis were noted. Epithelial cells forming keratin were demonstrated by a positive histochemical reaction with Rhodanile Blue. Immunohistochemical studies showed a positive reaction for CAM 5.2 indicating that the ROE‐2B cells express the cytokeratins of simple or glandular epithelia. The ROE‐2B cell line will be useful for studies on in vitro biological behaviour of odontogenic epithelial cells, and may allow the establishment of in vitro models of odonto