应用共聚焦成像评估甲氨蝶呤诱导的3D人肝脏类器官中的纤维化水平  

Application of Confocal Imaging to Detect Methotrexate-Induced Fibrosis in 3D Human Liver Organoids

作　　者：吴晓珊[1,2,3] 邓元荣 WU Xiao-shan;DENG Yuan-rong(School of Pharmacy,Fujian Health college,Fuzhou 350101,China;School of biotechnology,East China University of Science and Technology,Shanghai 200237,China;Shanghai Institute of Nutrition and Health,Shanghai 200031,China)

机构地区：[1]福建卫生职业技术学院药学院,福建福州350101 [2]华东理工大学生物工程学院,上海200237 [3]中国科学院上海营养与健康研究所,上海200031

出　　处：《海峡药学》2022年第11期32-35,共4页Strait Pharmaceutical Journal

基　　金：福建卫生职业技术学院应用技术协同创新研究面上项目(2018-2-9)。

摘　　要：目的 3D类器官培养的兴起为复杂疾病的模拟、药物筛选等方面提供了新型的解决方案,然而如何将传统的2D细胞的成像或检测技术应用于3D类器官,是目前研究人员亟待解决的一大问题。方法 采用免疫荧光法对甲氨蝶呤诱导的3D肝脏类器官中的纤维化蛋白CollagenⅠ进行染色。采用共聚焦显微镜进行成像并用Fiji软件对纤维化蛋白的成像结果进行分析定量。结果 采用10μM和30μM甲氨蝶呤处理7天的肝脏类器官与溶剂组相比呈现出显著的纤维化。结论 Z-stack共聚焦成像在保留类器官原有结构的基础上,为MTX诱导的3D肝类器官纤维化的量化,提供了一种非侵入性的、简便的方式。OBJECTIVE The rise of 3D organoid culture provides new solutions for the simulation of complex diseases or drug screening.However, how to translate traditional 2D cell imaging or detection technology to 3D organoids is an urgent problem for researchers.METHODS The immunofluorescence stained fibrotic protein-Collagen I in methotrexate-induced 3D liver organoids was detected by confocal imaging.The immunofluorescence of fibrotic protein was analyzed and quantified by Fiji software.RESULTS Liver organoids treated with 10 μM or 30 μM methotrexate for 7 days exhibited significant fibrosis compared to the vehicle group.CONCLUSION Z-stack confocal imaging provides a non-invasive and convenient way to quantify MTX-induced 3D hepatic organoid fibrosis.

关 键 词：3D肝脏类器官 共聚焦成像 甲氨蝶呤 肝纤维化 

分 类 号：R965[医药卫生—药理学]