Liver organoids were seeded and kept 7–10 days under the liver medium explained above (EM, expansion medium) supplemented with BMP7 (25 ng/ml). Then, the cultures were split and seeded accordingly in this EM supplemented with BMP7 for at least 2–4 days. Then, medium was changed to the differentiation medium (DM): AdDMEM/F12 medium supplemented with 1% N2 and 1% B27 and containing EGF (50 ng/ml), gastrin (10 nM, Sigma), HGF (25 ng/ml), FGF19 (100 ng/ml), A8301 (500 nM), DAPT (10 uM), BMP7 (25 ng/ml), and dexamethasone (30 uM). Differentiation medium was changed every 2–3 for a period of 11–13 days.
To assess hepatocyte function, culture medium was collected 24 hr after the last medium change. Functional studies were performed in the collected supernatant or in whole organoids, as described in theExtended Experimental Procedures .
To assess hepatocyte function, culture medium was collected 24 hr after the last medium change. Functional studies were performed in the collected supernatant or in whole organoids, as described in the
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