Journal: Stem Cell Research & Therapy

Article Title: Generation of human hepatobiliary organoids with a functional bile duct from chemically induced liver progenitor cells

doi: 10.1186/s13287-024-03877-z

Figure Lengend Snippet: The induction of HBO formation in an ultralow attachment dish by different induction media. A . Schematic representation of HBO induction. SHM?small hepatocyte medium, was described in the Methods section; FAC, SHM supplied with 10% FBS with A-83-01 and CHIR99021; BIM, BD induction medium; YAC, SHM medium with 10μM Y-27632, 0.5μM A-83-01, 3μM CHIR99021. B . The morphology of HBOs in YAC, BIM and FAC media. Scale bar = 200 μm. C . The diameters of the HBOs in different induction media were heterogeneous. The diameters of YAC-HBOs showed a high degree of variation. D . H&E staining of a cross-slide of an HBO, the HBOs showed a biliary lumen. Scale bar = 100 μm

Article Snippet: Four hours later, the culture medium was changed to small chemically reprogrammed culture medium, which was descripted as the reported that small hepatocyte medium (SHM) was DMEM/F12 containing 2.4 g/L NaHCO 3 and L-glutamine (Life Technologies, Tokyo, Japan) and supplemented with 5 mM HEPES, 30 mg/L L-proline, 0.05% BSA, 10 ng/mL EGF (all from Sigma‒Aldrich Japan, Tokyo, Japan), insulin-transferrin-serine (ITS)-X (Life Technologies, Tokyo, Japan), 10 − 7 M dexamethasone (Dex) (Fuji Pharma Co. Ltd., Tokyo, Japan), 10 mM nicotinamide (Sigma‒Aldrich, Tokyo, Japan), 1 mM ascorbic acid-2 phosphate (Wako Pure Chemical, Osaka, Japan), 100 U/mL penicillin, and 100 mg/mL streptomycin (Life Technologies, Tokyo, Japan) supplied with two small chemical molecules of 0.5 µM A-83-01 (TGF-b type I receptor inhibitor, Wako Pure Chemical, Tokyo, Japan), 3 µM CHIR99021 (GSK-3 inhibitor, AdooQ BioScience, CA, USA) and 10% FBS, which would be called FAC medium [ ].

Techniques: Staining