子宫内膜异位症在位内膜上皮类器官模型的建立及鉴定  

Establishment and identification of in-place endometrial epithelial organoids in endometriosis

作　　者：张瑞琪 杨玉娥 马远[1] 李博巍 李贝 哈春芳[2,3] Zhang Ruiqi;Yang Yu'e;Ma Yuan(Ningxia Medical University,Yinchuan 750004)

机构地区：[1]宁夏医科大学,银川750004 [2]宁夏医科大学总医院,银川750004 [3]宁夏医科大学生育力保持教育部重点实验室,银川750004

出　　处：《现代妇产科进展》2024年第6期427-432,共6页Progress in Obstetrics and Gynecology

基　　金：宁夏回族自治区自然科学基金项目(No:2023AAC03589)。

摘　　要：目的:探讨并验证构建人子宫内膜异位症(EMs)患者在位内膜上皮类器官模型的可行性及影响因素。方法:选择经腹腔镜手术并病理确诊为卵巢型EMs囊肿患者的新鲜在位内膜组织10例,组织消化法将在位内膜消化成单细胞,在基质胶Matrigel中培养7d构建EMs在位内膜上皮类器官模型。通过形态学、免疫组化、HE染色、透射电镜检测等,与原在位内膜组织对比进行模型鉴定;雌激素1、5、10nmol/L干预后观察在位内膜上皮类器官直径的变化,初步探讨雌激素在EMs在位内膜上皮类器官构建中的作用。结果:10例患者在位内膜组织均成功构建EMs在位内膜上皮类器官,光学显微镜下显示在位内膜上皮类器官的数量可观,形态上呈类圆形3D立体结构,周围可见腺体结构;免疫组化显示上皮源性标记物及雌孕激素受体表达阳性属于内膜源性上皮;HE染色显示在高倍镜下构建的在位内膜上皮类器官排列成腺腔样结构但腺体排列稀疏,较不规则,腔体内部分细胞形成分隔与原组织腺上皮结构相似;透射电镜显示在位内膜上皮类器官的内部超微结构与原在位内膜组织具有高度相似性;在培养基中加不同浓度雌二醇,10nmol/L雌激素浓度干预第6d的类器官直径明显大于对照组(P<0.05)。结论:体外成功构建的EMs在位内膜上皮类器官模型具有上皮性特征,与原在位内膜组织对比高度重现来源组织的结构、病理特性,可替代人子宫内膜异位症在位内膜组织用于EMs疾病药物的干预与基础实验的研究的建模。Objective:To explore and verify the feasibility and influencing factors of constructing in-situ endometrial epithelial organoid model in human endometriosis patients.Methods:10 cases of fresh in-situ endometrial tissue from patients with ovarian endometriosis cyst diagnosed by laparoscopy and pathological diagnosis were selected.The in-situ endometrial tissue was digested into single cells by tissue digestion method.The human endometriosis eutopic epithelial organoids were cultured in Matrigel for 7 days.The model was identified by morphology,immunohistochemistry,HE staining and transmission electron microscopy.The diameter changes of in-place endometrial epithelial organoids were observed after the intervention of estrogen at 1nmol/L,5nmol/L and 10nmol/L,and the role of estrogen in the construction of in-place endometrial epithelial organoids was initially discussed.Results:EMs endometrial organoids were successfully constructed in the endometrial tissues of 10 patients.Optical microscopy showed that there were a considerable number of endometrial organoids in the endometrial tissues,and their morphology was similar to circular 3D structure with glandular structures around them.Immunohistochemistry showed that epidermal markers and estrogen receptor expression were positive in endometrium-derived epithelium.HE staining showed that the on-site endometrial epithelial organoids constructed under high power microscope were arranged in a glandular structure,but the glands were arranged sparsely and irregularly,and some cells in the cavity formed comparttures similar to the glandular epithelium structure of the original tissue.Transmission electron microscopy(TEM)showed that the internal ultrastructure of in situ endometrial epithelial organoids was highly similar to that of the original in situ endometrial tissues.When different concentrations of estradiol were added to the medium,the organoid diameter was significantly larger than that of the control group on day 6 after the intervention of 10nmol/L estrogen concent

关 键 词：子宫内膜异位症 在位内膜腺上皮类器官 模型 雌激素 

分 类 号：R711.71[医药卫生—妇产科学]