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. 2022 Sep;192(9):1200–1217. doi: 10.1016/j.ajpath.2022.05.007

Figure 2.

Figure 2

CCA organoids with multiple cell lines. Three-dimensional (3D) CCA organoids include multiple cell lines, such as CCA cell lines (cholangiocytes; CHOL), liver sinusoidal endothelial cells (LSECs), and hepatic stellate cells (HSCs) (representative). Other liver cells, such as hepatocytes and Kupffer cells, can also be added, if needed. These organoids can be made scaffold-free without any biomaterial (eg, Matrigel) in a low-binding plate using culture mediums and can be kept for 14 to 28 days. Evidence suggests that CCA cells express cytokeratin (CK)-7, a cholangiocyte marker, and organoids can be stained with other cellular markers (eg, vascular endothelial cadherin; VE-Cad) that are present in organoids. CCA organoids can be exposed to immune cells, as seen at the bottom right. Bright-field microscopy shows a 3D CCA organoid exposed to mast cells. Scale bar = 500 μm. Original magnification, ×10.