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International Journal of Bioprinting Liver printing: from structure to application

Figure 9. Biomanufacturing of bile ducts. (A) Biliary tree, bile duct, cholangiocytes, and ductal bile formation. Adapted with permission from Wang
et al. (B) Cholangiocytes encapsulated in the liver decellularized extracellular matrix (dECM) spontaneously form duct-like structures. Adapted with
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permission from Lewis et al. (C) Immunofluorescence images of primary cholangiocytes illustrate the differential expression of representative region
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markers. Scale bars: 50 μm. Adapted with permission from Sampaziotis et al. (D) Hepatobiliary connections in co-culture optical cross-sections along
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broken lines in the left panel are displayed in the right panels. The BC within the HNF4α+ hepatocyte clusters (arrowheads in upper right panels) connect
to the lumen, which consists of CK19+ cholangiocytes and HNF4α+ hepatocytes (arrowheads in middle right panels), and then eventually reach the duct
of CK19+ cholangiocytes (arrowheads in lower right panels). At 4 weeks after MG overlay, the immunostaining with phalloidin (white), anti-HNF4α
antibody (red), anti-CK19 antibody (green), and Hoechst 33342 (blue) was repeated five times independently. Three fields were examined in each sample.
Three out of 15 areas were further used to collect serial optical sections for 3D reconstruction. The representative images are displayed in this figure. Scale
bars: 50 μm. Adapted with permission from Tanimizu et al. (E) Cell-printing of the 3D liver-on-a-chip with vascular/biliary systems. (F) The evaluation
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of cell positioning after bioprinting 3D liver-on-a-chip. Scale bar: 100 μm. (G) Cell-printed 3D liver-on-a-chip with dual fluidic condition. Adapted with
permission from Lee et al. Abbreviations: BC, bile canaliculi; CBD, common bile duct; CK19: Cytokeratin 19; DAPI, 4’,6-diamidino-2-phenylindole; GB,
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gallbladder; HNF4α: Hepatocyte Nuclear Factor 4 alpha; HUVECs, human umbilical vein endothelial cells; IHD, intrahepatic ducts; Krt19, Keratin 19;
MG: Matrigel; Sox17, SRY (sex-determining region Y)-box 17; Sox4, SRY (sex-determining region Y)-box 4; TFF2, Trefoil Factor 2.
Volume 10 Issue 5 (2024) 141 doi: 10.36922/ijb.3819