人源胎盘部位滋养细胞肿瘤类器官模型的构建与鉴定  

The construction and identification of adult-derived placental site trophoblastic tumor organoid

作　　者：张赛 周佳宜[1] 吴静 于焕镝 丁雨潇 杜琰[2] 鹿欣 赵洪波[1] ZHANG Sai;ZHOU Jia-yi;WU Jing;YU Huan-di;DING Yu-xiao;DU Yan;LU Xin;ZHAO Hong-bo(Biobank,Obstetrics and Gynecology Hospital,Fudan University,Shanghai 200011,China;Research Center for Clinical Medicine,Obstetrics and Gynecology Hospital,Fudan University,Shanghai 200011,China;Department of Gynecology,Obstetrics and Gynecology Hospital,Fudan University,Shanghai 200011,China)

机构地区：[1]复旦大学附属妇产科医院生物样本库,上海200011 [2]复旦大学附属妇产科医院临床研究中心,上海200011 [3]复旦大学附属妇产科医院妇科,上海200011

出　　处：《复旦学报（医学版）》2024年第5期800-806,共7页Fudan University Journal of Medical Sciences

基　　金：国家重点研发计划(2023YFC2705801);上海市卫健委卫生行业临床研究专项青年项目(20234Y0067);国家自然科学基金(82371687)。

摘　　要：目的构建并鉴定人源胎盘部位滋养细胞肿瘤(placental site trophoblastic tumor,PSTT)的类器官模型。方法选取PSTT组织进行消化和分离,得到肿瘤细胞后进行基质胶包埋,利用特定的类器官培养基进行培养。HE染色观察类器官模型的组织学形态,通过免疫组化和免疫荧光染色分析中间型滋养细胞标志物人胎盘催乳素(human placental prolactin,HPL)、人类白细胞抗原-G(human leukocyte antigen-G,HLA-G)和胎盘碱性磷酸酶(placental alkaline phosphatase,PLAP)的表达,鉴定构建的PSTT类器官模型与PSTT组织之间的一致性。同时对构建的模型进行传代及冻存、复苏,显微镜下分析其生长形态与速度,探讨其作为类器官模型在PSTT基础及临床转化研究应用的可能性。结果构建的类器官模型可稳定增殖并传代,可见其从小微球状态长成紧致实性球体或带有滤泡样结构的球体,约7~10天可长满并传代,冻存及复苏后细胞状态稳定;经HE染色发现类器官中细胞形态与PSTT原代肿瘤细胞相似;免疫组化与免疫荧光染色鉴定结果显示,PSTT类器官组织高表达中间型滋养细胞的标志分子HPL、HLA-G和PALP,提示构建的类器官模型主要包含中间型滋养细胞。结论成功建立了人源PSTT类器官(adult-derived PSTT organoid,ADPO)模型并进行鉴定。Objective To construct and identify an organoid model of human placental site trophoblastic tumor(PSTT).Methods The tumor cells were obtained by digesting and separating the PSTT tissues and then embedded in Matrigel.The organoids were cultured in the specific organoid medium.The histological morphology of the organoid model was observed by HE staining and the expression levels of the PSTT specific markers[human placental prolactin(HPL),human leukocyte antigen-G(HLA-G)and placental alkaline phosphatase(PLAP)]were detected by immunohistochemistry and immunofluorescence,so as to evaluate the consistency between the organoid model and the PSTT tissue.Meanwhile,the morphology and forming efficiency of the constructed model were observed under a microscope after primary culture,passage generation and cryopreservation to evaluate its potential application as an organoid model in basic and clinical translational research of PSTT.Results The constructed organoid model could proliferate stably,growing from small microspheres into compact solid spheres or spheres with follicle-like structures,and could passage after fully grown in 7-10 days.The cell state remained stable after passage,frozen storage and recovery.HE staining showed that the morphology of the cells in the organoids was similar to that of the primary PSTT tumor cells,and immunofluorescence staining showed that the organoids highly expressed HLA-G and lowly expressedβ-HCG,indicating that the constructed organoid model mainly contained intermediate trophoblast.Conclusion The adult-derived PSTT organoid(ADPO)models were successfully established.

关 键 词：类器官 胎盘部位滋养细胞肿瘤(PSTT) 细胞培养 

分 类 号：R711.74[医药卫生—妇产科学]