目的：建立腮腺多形性腺瘤（PAs）患者来源的类器官模型，并初步表征其组织学、相关生物标志物和功能。方法：采集口腔颌面部外科手术中的新鲜肿瘤组织标本。将采集的组织在头颈部肿瘤类器官培养系统中进行处理和培养，成功建立了4例腮腺多形性腺瘤的类器官模型。通过光学显微镜记录 PA 类器官的体外生长。将成功建立的类器官进行传代和冷冻保存，并对冷冻保存的PA类器官进行复苏和再培养，观察其活力和类器官再生能力。对类器官进行组织学表征以及相关标记物和功能蛋白的表征和检测，并将其与患者来源的组织进行比较。结果：构建的多形性腺瘤类器官模型呈现致密、致密的三维球形结构。苏木精和伊红染色表明类器官与其来源组织之间的形态学相似性。免疫组织化学显示类器官和来源肿瘤组织中钙调蛋白、CK7 和 EMA 细胞质染色呈阳性，表明类器官与其来源组织之间具有一致的组织病理学特征。类器官过碘酸希夫染色显示糖原呈阳性染色，阳性染色位于类器官的内部和外围，表明类器官具有与唾液腺一样的分泌功能。结论：我们成功构建了源自患者样本的多形性腺瘤类器官。该模型忠实地复制了源组织的组织形态和生物标志物，并表现出与粘液分泌相关的生物学功能。它是研究唾液腺肿瘤发生和进展的有价值的体外模型。

Objective: To establish patient-derived organoid models of pleomorphic adenomas (PAs) of the parotid gland and preliminarily characterize their histology, related biomarkers and functions. Methods: Fresh tumor tissue specimens were collected from surgical procedures of Oral and Maxillofacial Department. The harvested tissues were processed and cultured in a head and neck tumor organoid culture system, resulting in the successful establishment of organoid models from four cases of parotid gland pleomorphic adenomas. The in vitro growth of PA organoids was recorded by light microscopy. The successfully established organoids were passaged and cryopreserved, and the cryopreserved PA organoids were revived and re-cultured to observe their viability and organoid regeneration ability. Histological characterization, as well as characterization and detection of related markers and functional proteins, were performed on the organoids, comparing them with the patient-derived tissues. Results: The constructed organoid model of pleomorphic adenoma exhibited a dense and compact three-dimensional spherical structure. Hematoxylin and eosin staining indicated morphological similarities between the organoid and its tissue of origin. Immunohistochemistry showed positive cytoplasmic staining for Calponin, CK7, and EMA in both the organoid and the source tumor tissue, suggesting consistent histopathological characteristics between the organoid and its tissue of origin. Periodic acid-Schiff staining of the organoid showed positive staining for glycogen, with positive staining located in the interior and periphery of the organoid, indicating that the organoid possessed secretory functions like the salivary gland. Conclusion: We successfully constructed organoids of pleomorphic adenoma derived from patient samples. This model faithfully replicates the tissue morphology and biomarkers of the source tissue and exhibits biological functions associated with mucus secretion. It serves as a valuable in vitro model for studying the development and progression of salivary gland tumors.