球体
肝星状细胞
细胞
细胞培养
生物
病理
三维细胞培养
电池类型
体外
免疫荧光
细胞生物学
多细胞生物
细胞外基质
染色
免疫学
抗体
医学
生物化学
遗传学
作者
M. Couteau,Lynda Aoudjehane
出处
期刊:Methods in molecular biology
日期:2024-01-01
卷期号:: 143-152
标识
DOI:10.1007/978-1-0716-3694-7_11
摘要
In vitro studies on liver diseases, such as non-alcoholic fatty liver disease, fibrosis, and hepatocellular carcinoma, are traditionally performed in two-dimensional (2D) cultures of isolated primary cells or immortalized cell lines. However, this approach has limitations, as 2D cultures inadequately replicate the cell–cell and cell–extracellular matrix interactions found in three-dimensional (3D) environments. To overcome this limitation, various 3D models, such as spheroids, have been developed. These spheroids serve as simplified biomimetic in vitro models for studying liver diseases. They can be generated using a variety of cells from healthy and pathological tissues, including liver cancer. Here, we present a comprehensive protocol for performing immunofluorescent staining and confocal imaging on whole human hepatic multicellular spheroids, utilizing primary cells or cell lines. The immunofluorescence technique is a potent tool to understand the spatial distribution of different cell types within the spheroids and define the interactions that occur among these cells.
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