Isolation, Culture, and Characterization of Primary Salivary Gland Cells

Primary cells are an essential tool for in vitro studies and are obtained directly from living tissues or organs. They closely mimic the physiological state and maintain in vivo functions for short periods of time under optimal conditions. Isolation and culture of salivary gland (SG) cells are useful to decipher the various mechanisms involved in salivary gland dysfunction. However, unlike some other primary cell cultures, SG cell cultures from patient-derived tissues present several challenges. They are difficult to obtain, culture, expand, and characterize due to their sensitive heterogenous cell population and limited expansion potential. In addition, the majority of saliva-secreting acinar cells fail to maintain a differentiated state ex vivo for long periods, and eventually succumb to an acinar-to-ductal metaplasia, losing their secretory phenotype and functions. Herein, we describe two detailed protocols for primary SG cell isolation, culture, and expansion from human (or mouse) salivary tissues using serum-free culture media. We also describe the growth kinetics of these primary cells along with their immunocytochemical characterization. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Preparation of SG single-cell culture from freshly obtained human or mouse SG tissues. Basic Protocol 2: Preparation of SG explant culture from freshly obtained human or mouse SG tissues.