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.
Highlights
Development of restorative therapies for salivary gland dysfunction has been significantly hampered by a lack of accessible and pertinent cell culture models
Salivary glands are made of several cell types, including the saliva-producing acinar cells, ductal cells modifying the saliva as it travels through the lumen, and myoepithelial cells mediating acinus contraction
At late passages immortalized salivary gland epithelial cells (iSGECs)-pSS1 gave rise to colonies appearing that were mixed in size, with polygonal or cobblestone-like cells with differing rates of cytoplasmic complexity (Figure 1A). iSGEC-nSS1 late passages demonstrated a mix of small polygonal and filiform-appearing cells (Figure 1B)
Summary
Development of restorative therapies for salivary gland dysfunction has been significantly hampered by a lack of accessible and pertinent cell culture models. Cell culture models for pSS and xerostomia have been limited, leading researchers to rely on cancer cell lines or short-term cultures of primary salivary gland epithelial cells (SGECs) [4,5]. The cell lines extensively used in salivary research (e.g., HSG, A253, NS-SV-AC) are of male origin [4,6,7,8]. These cell lines may not recapitulate the pathological processes occurring in pSS, a disease with a higher incidence in female patients [9]
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
