患者来源腮腺多形性腺瘤类器官模型构建和组织学与功能初步表征  

Patient-derived parotid gland pleomorphic adenoma organoid model and its preliminary histological and functional characterization

作　　者：陈雨鑫 王若雨 王秋旭 廉瑞青 王玉娥 梁珊珊 Chen Yuxin;Wang Ruoyu;Wang Qiuxu;Lian Ruiqing;Wang Yu'e;Liang Shanshan(Liaoning Provincal Key Laboratory of High-Throughput Screening or Targeted Drug Transformation of Molecular Markers of Breast and Digestive Tumors,Dalian 116001,China;Department of Oral and Maxillofacial Surgery,Affiliated Zhongshan Hospital of Dalian University,Dalian 116001,China;Department of Pathology,Affiliated Zhongshan Hospital of Dalian University,Dalian 116001,China)

机构地区：[1]辽宁省乳腺及消化肿瘤分子标志物高通量筛选及靶向药物转化重点实验室,大连116001 [2]大连大学附属中山医院口腔颌面外科,大连116001 [3]大连大学附属中山医院病理科,大连116001

出　　处：《中华口腔医学杂志》2024年第6期604-609,共6页Chinese Journal of Stomatology

基　　金：国家自然科学基金(82172822);大连市科技局高层次人才创新支持计划-尖端和领军人才项目(2021RD02)。

摘　　要：目的构建患者来源腮腺多形性腺瘤(PA)类器官模型,并对其组织学、相关标志物和功能进行初步表征。方法收集大连大学附属中山医院口腔颌面外科术中取材新鲜肿瘤组织标本,采用头颈部肿瘤类器官培养体系进行体外培养,构建腮腺PA类器官。观察PA类器官的体外生长情况并进行光镜拍摄。对培养成功的类器官进行传代和冻存,并对冻存的PA类器官进行复苏再培养,观测其活性和PA类器官再形成能力。对比患者来源组织,对PA类器官进行组织学表征,相关标志和功能蛋白进行表征与检测,评价PA类器官对来源组织的复现性。结果构建的腮腺PA类器官形成紧实致密球状结构,HE染色提示类器官与其来源组织形态相似,免疫组化可见类器官及其来源肿瘤组织钙调蛋白、细胞角蛋白7、上皮膜抗原胞质阳性,提示类器官与其来源组织的病理学特征一致;PA类器官过碘酸希夫(PAS)染色及淀粉酶消化后PAS染色呈阳性,阳性着色位于类器官球体内部及外周,提示PA类器官具有类似于腮腺的分泌功能。结论成功构建患者来源PA类器官,该模型在组织形态和标志物上可高度复现来源组织情况,并具有黏液分泌的生物学功能,是研究腮腺肿物发生发展的良好体外模型。ObjectiveTo establish patient-derived organoid models of pleomorphic adenomas(PA)of the parotid gland and preliminarily characterize their histology,related biomarkers and functions.MethodsFresh 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 to establish organoid models from 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.ResultsThe 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,cytokeratin 7,and epithelial membrane antigen 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.ConclusionsThis study 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 tu

关 键 词：类器官 腺瘤 多形性 体外三维培养 组织学表征 肿瘤异型性 

分 类 号：R739.87[医药卫生—肿瘤]