Establishment of a convenient system for the long‐term culture and study of non‐neoplastic human salivary gland epithelial cells

Abstract

Epithelial cells appear to play an important role in the initiation and maintenance of autoimmune lesions in the salivary glands of patients with Sjogren's syndrome. Therefore, the detailed study of immunological function of salivary gland epithelial cells (SGEC) may provide useful information for the understanding of Sjögren's syndrome pathogenesis. In this report we aimed to formulate a protocol for the establishment of human non-neoplastic SGEC lines as a tool for the study of the physiology and pathophysiology of these cells. Pointing towards a practical approach, we sought to establish SGEC lines from quite a limited amount of biopsy tissue obtained during the diagnostic evaluation of patients. Herein, the favorable conditions for the long-term maintenance of human non-neoplastic SGEC lines are presented and involve the successive application of a serum-containing and a serum-free culture medium, supplemented with essential epithelial growth factors. This protocol has been found reliable and convenient, as attested by the reproducible establishment of non-neoplastic SGEC lines. The analysis of SGEC phenotypic features, as well as a coculture system for the study of interactions between epithelial cells and lymphocytes, are also described. Such techniques may provide valuable means for the functional and molecular investigation of human SGEC and particularly for the study of Sjögren's syndrome and other disorders of glandular epithelia.

B‐cell‐activating factor expressions in salivary epithelial cells after dsRNA virus infection depends on RNA‐activated protein kinase activation

“…Primary cultures of SGEC were established from minor salivary glands as described [35]. Briefly, each lobule was cut into small fragments and set in six 75-cm 2 flasks with basal epithelial medium (a 3:1 mixture of Ham's F-12 and DMEM) supplemented with 2.5% FBS, EGF (10 ng/mL), hydrocortisone (0.4 mg/mL), insulin (0.5 mg/ mL), penicillin (100 IU/mL) and streptomycin (100 mg/mL) and incubated at 371C with 5% CO 2 .…”

Establishment of Functional Acinar-like Cultures from Human Salivary Glands

“…Few studies have reported the isolation and growth of primary epithelial cells from explants of salivary gland tissues (Sens et al 1985;Sabatini et al 1991;Okura et al 1993;Dimitriou et al 2002). However, maintenance of the acinar phenotype for long periods of time and preservation of the acinar cell functionality have not been demonstrated yet (Redman and Quissell 1993).…”

A Novel B7-2 (CD86) Splice Variant with a Putative Negative Regulatory Role

“…The purity and epithelial origin of cultured SGEC lines was routinely verified by morphology, the uniform expression of epithelial-specific markers and the absence of markers indicative of lymphoid/monocytoid cells (16). Distinct SGEC lines have not been found to differ in morphological features, proliferation rate or survival (16).…”

“…Non-neoplastic, long-term cultured SGEC lines and salivary gland fibroblasts were established from minor salivary gland biopsy samples obtained with informed consent from patients undergoing diagnostic evaluation for possible Sjögren's syndrome, as previously described (16). The purity and epithelial origin of cultured SGEC lines was routinely verified by morphology, the uniform expression of epithelial-specific markers and the absence of markers indicative of lymphoid/monocytoid cells (16).…”