Rho-associated kinase inhibitor enhances the culture condition of isolated mouse salivary gland cells in vitro

•We describe here an advanced culture condition for isolated mouse salivary gland cells in vitro.•Normally, culture and maintaining characteristics of isolated salivary gland cells are hampered.•Therefore, this culture condition may serve as an efficient tool for culturing salivary gland cells for their application. Hyposalivation because of curative radiation therapy in patients with head and neck cancer is a major concern. At present, there is no effective treatment for hyposalivation, highlighting the importance of cell therapy as a new therapeutic approach. To provide functional cells for cell replacement therapy, it is important to overcome the limitations of current in vitro culture methods for isolated salivary gland cells. Here, we suggest an improved culture condition method for the cultivation of isolated salivary gland cells. The dissociated submandibular salivary gland cells of mice were seeded and treated with Rho-associated kinase (ROCK) inhibitor (Y-27632), which resulted in an increase in their cell adhesion, viability, migration, and proliferation. In particular, ROCK inhibitor treatment maintained the expression of α-amylase in the primary cultured salivary gland cells for a long time as compared with untreated cells. The expression of C-Met, a ductal cell marker, was increased in cells treated with ROCK inhibitor. This modified culture condition may serve as an easy and convenient tool for culturing primary salivary gland cells for their application in hyposalivation therapy.