In vitro culture of human salivary gland epithelial cells (SGEC) is still a challenge. A high quantity and quality of cells are needed for the cultivation of 3D matrices. Furthermore, it is known that DNA damage is supposed to be an important factor involved in carcinogenesis. This study investigates cellular function and DNA integrity of human SGEC during 3 passage steps in 2 groups (group 1: n = 10; group 2: n = 9). Cellular function was analyzed by immunofluorescence, transmission electron microscopy (TEM), and quantitative real-time polymerase chain reaction (qPCR). DNA integrity was tested via the comet assay. Immunohistochemistry and qPCR results showed stable α-amylase and pan-cytokeratin levels; TEM revealed functional cells; and no significant DNA damage could be detected in the comet assay during 3 culture steps. The study shows that not only at cellular but also at DNA level human SGEC can be safely quantified over 3 passages for preclinical tissue engineering without loss of differentiation and function.

中文翻译：

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人唾液腺上皮细胞 2D 体外培养过程中细胞功能和 DNA 完整性的研究

人唾液腺上皮细胞 (SGEC) 的体外培养仍然是一个挑战。培养 3D 矩阵需要大量和高质量的细胞。此外，已知DNA损伤被认为是参与致癌作用的重要因素。本研究调查了人类 SGEC 在 3 个传代步骤中的细胞功能和 DNA 完整性，分为 2 组（第 1 组：n = 10；第 2 组：n = 9）。通过免疫荧光、透射电子显微镜 (TEM) 和定量实时聚合酶链反应 (qPCR) 分析细胞功能。DNA完整性通过彗星试验进行测试。免疫组织化学和 qPCR 结果显示稳定的 α-淀粉酶和泛细胞角蛋白水平；TEM 显示功能细胞；在彗星试验中，在 3 个培养步骤中未检测到明显的 DNA 损伤。