**Braxton D. Noll, Alexandre Grdzelishvili, Michael T. Brennan, Farah Bahrani Mougeot and Jean-Luc C. Mougeot \***

Carolinas Medical Center-Atrium Health, Charlotte, NC 28203, USA; braxton.noll@atriumhealth.org (B.D.N.); alexandre.grdzelishvili@atriumhealth.org (A.G.); mike.brennan@atriumhealth.org (M.T.B.); farah.mougeot@atriumhealth.org (F.B.M.)

**\*** Correspondence: jean-luc.mougeot@atriumhealth.org

Received: 30 September 2020; Accepted: 23 November 2020; Published: 25 November 2020

**Abstract:** Primary Sjögren's Syndrome (pSS) is an autoimmune disease mainly affecting salivary and lacrimal glands. Previous pSS studies have relied on primary cell culture models or cancer cell lines with limited relevance to the disease. Our objective was to generate and characterize immortalized salivary gland epithelial cells (iSGECs) derived from labial salivary gland (LSG) biopsies of pSS patients (focus score > 1) and non-Sjögren's Syndrome (nSS) xerostomic (i.e., sicca) female patients. To characterize iSGECs (*n* = 3), mRNA expression of specific epithelial and acinar cell markers was quantified by qRT-PCR. Protein expression of characterization markers was determined by immunocytochemistry and Western blot. Secretion of α-amylase by iSGECs was confirmed through colorimetric activity assay. Spheroid formation and associated alterations in expression markers were determined using matrigel-coated cell culture plates. Consistent mRNA and protein expressions of both epithelial and pro-acinar cell markers were observed in all three iSGEC lines. When cultured on matrigel medium, iSGECs formed spheroids, secreted α-amylase after β-adrenergic stimulation, and expressed multiple acinar cell markers at late passages. One iSGEC line retained adequate cell morphology without a loss of SV40Lt expression and proliferation potential after over 100 passages. In conclusion, our established iSGEC lines represent a viable model for salivary research due to their passaging capacity and maintenance of pro-acinar cell characteristics.

**Keywords:** Sjögren's syndrome; xerostomia; salivary gland; SGEC; immortalization; acinar; ductal; spheroid
