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Label-retaining Cells in the Rat Submandibular Gland

Masaya Kimoto, Yoshiaki Yura, Mitsunobu Kishino, Satoru Toyosawa and Yuzo Ogawa

Department of Oral Pathology (MKimoto,MKishino,ST,YO) and Department of Oral and Maxillofacial Surgery II (MKimoto,YY), Osaka University Graduate School of Dentistry, Osaka, Japan

Correspondence to: Yuzo Ogawa, DDS, PhD, Department of Oral Pathology, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka 565-0871, Japan. E-mail: ogawa{at}dent.osaka-u.ac.jp

To identify stem cells in salivary glands, label-retaining cells (LRCs) were established in rat submandibular glands. Developing and regenerating glands were labeled with bromodeoxyuridine (BrdU). To cause gland regeneration, the glands were injured by duct obstruction. BrdU LRCs were observed in all the parenchymal structures except for the acinus of the glands labeled during regeneration. Among these LRCs, a few, but not many, expressed neither keratin18 (K18; an acinar/duct cell marker) nor -smooth muscle actin (SMA; a myoepithelial cell marker), and thus were putative stem cells. These (K18 and SMA)^neg LRCs were invariably observed in the intercalated duct and the excretory duct. In the intercalated duct, they were at the proximal end bordering the acinus (the neck of the intercalated duct). Next, to test the above identification, gland extirpation experiments were performed. LRCs were established by labeling developing glands with iododeoxyuridine (IdU) in place of BrdU. Removal of one submandibular gland forced the IdU-LRCs in the remaining gland to divide. They were labeled with chlorodeoxyuridine (CldU). The (K18 and SMA)^neg LRCs in the neck of the intercalated duct and in the excretory duct did not change in number or in IdU label. The CldU label appeared in these cells and then disappeared. These results indicate that the (K18 and SMA)^neg LRCs have divided asymmetrically and are thus considered salivary gland stem cells. (J Histochem Cytochem 56:15–24, 2008)

Key Words: stem cell • salivary gland • rat • immunohistochemistry

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