Tran S D, Wang J, Bandyopadhyay B C, Redman R S, Dutra A, Pak E, Swaim W D, Gerstenhaber J A, Bryant J M, Zheng C, Goldsmith C M, Kok M R, Wellner R B, Baum B J
National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland 20892, USA.
Tissue Eng. 2005 Jan-Feb;11(1-2):172-81. doi: 10.1089/ten.2005.11.172.
Therapeutic irradiation for head and neck cancer, and the autoimmune disease Sjogren's syndrome, lead to loss of salivary parenchyma. They are the two main causes of irreversible salivary gland hypofunction. Such patients cannot produce adequate levels of saliva, leading to considerable morbidity. We are working to develop an artificial salivary gland for such patients. A major problem in this endeavor has been the difficulty in obtaining a suitable autologous cellular component. This article describes a method of culturing and expanding primary salivary cells obtained from human submandibular glands (huSMGs) that is serum free and yields cells that are epithelial in nature. These include morphological (light and transmission electron microscopy [TEM]), protein expression (immunologically positive for ZO-1, claudin-1, and E-cadherin), and functional evidence. Under confocal microscopy, huSMG cells show polarization and appropriately localize tight junction proteins. TEM micrographs show an absence of dense core granules, but confirm the presence of tight and intermediate junctions and desmosomes between the cells. Functional assays showed that huSMG cells have high transepithelial electrical resistance and low rates of paracellular fluid movement. Additionally, huSMG cells show a normal karyotype without any morphological or numerical abnormalities, and most closely resemble striated and excretory duct cells in appearance. We conclude that this culture method for obtaining autologous human salivary cells should be useful in developing an artificial salivary gland.
头颈部癌症的放射治疗以及自身免疫性疾病干燥综合征会导致唾液腺实质丧失。它们是不可逆唾液腺功能减退的两个主要原因。这类患者无法产生足够的唾液，从而导致相当大的发病率。我们正在努力为这类患者开发一种人工唾液腺。这项工作中的一个主要问题是难以获得合适的自体细胞成分。本文描述了一种培养和扩增从人下颌下腺（huSMG）获得的原代唾液细胞的方法，该方法无血清且能产生本质上为上皮细胞的细胞。这些证据包括形态学（光学显微镜和透射电子显微镜 [TEM]）、蛋白质表达（对ZO-1、claudin-1和E-钙黏蛋白免疫阳性）以及功能方面的证据。在共聚焦显微镜下，huSMG细胞呈现极化并使紧密连接蛋白定位适当。TEM显微照片显示不存在致密核心颗粒，但证实了细胞间紧密连接、中间连接和桥粒的存在。功能测定表明，huSMG细胞具有高跨上皮电阻和低细胞旁液体移动速率。此外，huSMG细胞显示出正常的核型，没有任何形态或数量异常，并且在外观上最类似于纹状管和排泄管细胞。我们得出结论，这种获取自体人唾液细胞的培养方法在开发人工唾液腺方面应该是有用的。
