Figure 2

Characterization of 3D MCSs. (a) CCA and cholangiocyte cell lines were varied with seeding numbers (1,000 2,500 5,000 10,000 25,000 and 50,000 cells per well) and cultured for 48 h to observe the MCS aggregation under an inverted microscope. Scale bar represents 500 µm. (b) The 3D MCSs were embedded, sectioned, and stained with H&E. Scale bar represents 100 µm and 50 µm for zoom-in pictures. (c) The 3D MCSs exhibit typical zonation of hypoxia and oxidative stress determined by nuclei (DAPI, blue), Hypoxia probe (red), Oxidative stress probe (green). Scale bar represents 800 µm.