Salivary gland stem/progenitor cells are thought to be present in intercalated ductal cells, but the fact is unclear, In this Study, we sought to clarify if stem/progenitor cells are present in submandibular glands using colony assay, which is one of the stem cell assay methods. Using a low-density culture of submandibular gland cells of neonatal rats. we developed,I novel culture system that promotes single cell colony formation. Average doubling time for the colony-forming cells was 24.7 (SD = +/- 7.02) h, indicating high proliferative potency. When epidermal growth factor (EGF) and hepatocyte growth factor (HGF) were added to the medium. the number of clonal colonies increased greater than those cultured without growth factors (13.2 +/- 4.18 vs. 4.5 +/-1.73). The RT-PCR and immunostaining demonstrated expressing acinar, ductal, and myoepithelial cell lineage markers, This study demonstrated the presence of the salivary gland stem/progenitor cells that are highly proliferative and multipotent ill salivary glands. (c) 2005 Elsevier Inc. All rights reserved.