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Cellosaurus publication CLPUB00653

Publication number CLPUB00653
Authors Lin L.-C.
Title A new tool in human salivary gland research: the SMG-hu-1 cell line.
Citation Thesis MDSc (2017), McGill University Montreal, Canada
Web pages https://escholarship.mcgill.ca/concern/theses/9w0325503
Abstract Normal salivary processes are vital to the health of a patient. The salivary fluid has important functions such as digestion, lubrification, antibacterial protection and tooth remineralization. Salivary glands dysfunction such as hyposalivation can be observed in patients undergoing radiotherapy for head and neck cancer, and in patients suffering from Sjogren's syndrome, an autoimmune disease. Symptoms of hyposalivation include xerostomia (dry mouth), oral infections, tooth decay, pain, dysgeusia (distortion of taste) and disrupted speech. Numerous drugs and cell therapies have been proposed to reverse the impaired glandular secretion. Among the tools used for such research is the HSG cell line, commonly used in salivary gland research and mistakenly labeled as a submandibular ductal cell cell line. ATCC has recently released a list of cell lines cross-contaminated by HeLa, that included HSG. Despite the notice, some salivary gland research labs still use HSG. In this work, we describe the methodology for authenticating cell lines, assess the validity of previous HSG findings and develop a new salivary gland cell line. Objectives-We examine the authenticity and purity of two samples of HSG obtained from two independent labs using STR analysis. A literature review on HeLa cells is conducted to determine the validity of using HeLa- derived cells, namely HSG, as a model for salivary gland cells to support past salivary gland research findings that resulted from HSG. Lastly, a new human submandibular salivary gland cell line will be established and characterized. Results: STR analysis: HSG-Tran and HSG-Delporte have > 0.80 match with HeLa in DSMZ database. This confirms that both cell lines share a common ancestry (host) with HeLa. Literature review-based on reports of the karyotyping, cellular markers and membrane barrier function, HSG (HeLa) is not an appropriate model of normal human salivary gland acinar/ducal cells. Due to advanced mutations, it is even fair to say that HeLa is no longer a valid representation of human biology. Cell line establishment-The morphology of the SV40LT transfected salivary gland cells (SMG-hu-1) is highly similar to the non-transfected cells, both with an epithelial-like cobblestone morphology. SMG-hu-1 cells proliferate steadily (passage 10+) whereas non-transfected cells enter senescence after passage 1. Under immunofluorescence, SMG-hu-1 cells are positive for epithelial markers E-cadherin and cytokeratin. In turn, they are negative for vimentin (mesenchymal), CD31 (endothelial), alpha-smooth muscle actin (myoepithelial). AQP5 (serous acinar) staining is inconclusive. RNA expression reveals CK5, AQP5 and SV40. CFTR and CK19 expression are not found. Conclusions: STR analysis reveals that HSG is a HeLa derived cell line. Furthermore, as genotyping of the original cell line was not performed during its establishment, it will be impossible to authenticate a cell line that is supposedly an uncontaminated sample of HSG. Similarly, based on its characteristics and properties, HeLa derived cells, such as HSG, are not an accurate model of salivary gland cells. The use of HSG in salivary gland research should be discontinued and past findings should be reassessed with an accurate model, such as our newly established human submandibular salivary gland cell line (SMG-hu-1). SMG-hu-1 cells share the epithelial morphology of its non-transfected counterpart, but with significantly higher replicative capabilities. Immunofluorescence reveals presence of E-cadherin and cytokeratin. PCR and RT-PCR reveals expression of CK5, AQP5 and SV40. By utilizing this cell line, we look to further investigate salivary processes in hope to develop a cure to hyposalivation.
Cell lines CVCL_2517; HSG