An in vitro model of polycystic liver disease using genome-edited human inducible pluripotent stem cells

• Published in: Stem cell research 2018-10, Vol.32, p.17-24
• Main Authors: Kamiya, Akihide, Chikada, Hiromi, Ida, Kinuyo, Ando, Emi, Tsuruya, Kota, Kagawa, Tatehiro, Inagaki, Yutaka
• Format: Article
• Language: English
• Subjects: AC133 Antigen - metabolism; CD13 Antigens - metabolism; Cell Differentiation; Cell Line; Cells, Cultured; Cholangiocyte; Cysts - metabolism; Cysts - pathology; Flow Cytometry; Gene Editing - methods; Genome editing; Glucosidases - deficiency; Glucosidases - genetics; Human iPS cells; Humans; Intracellular Signaling Peptides and Proteins - deficiency; Intracellular Signaling Peptides and Proteins - genetics; Liver - cytology; Liver Diseases - metabolism; Liver Diseases - pathology; Polycystic liver disease
• ISSN: 1873-5061; 1876-7753
• DOI: 10.1016/j.scr.2018.08.018

In the developing liver, bile duct structure is formed through differentiation of hepatic progenitor cells (HPC) into cholangiocytes. A subtype of polycystic liver diseases characterized by uncontrolled expansion of bile ductal cells is caused by genetic abnormalities such as in that of protein kinase C substrate 80 K-H (PRKCSH). In this study, we aimed to mimic the disease process in vitro by genome editing of the PRKCSH locus in human inducible pluripotent stem (iPS) cells. A proportion of cultured human iPS cell-derived CD13+CD133+ HPC differentiated into CD13− cells. During the subsequent gel embedding culture, CD13− cells formed bile ductal marker-positive cystic structures with the polarity of epithelial cells. A deletion of PRKCSH gene increased expression of cholangiocytic transcription factors in CD13− cells and the number of cholangiocytic cyst structure. These results suggest that PRKCSH deficiency promotes the differentiation of HPC-derived cholangiocytes, providing a good in vitro model to analyze the molecular mechanisms underlying polycystic diseases. •The CD13− cultured hepatic progenitor cells derived from human iPS cells contains the phenotypes of cholangiocyte cells.•Using CRISPR/Cas9, PRKCSH-mutated human iPS cells related to polycystic liver disease were established.•PRKCSH deficiency promotes the differentiation of HPC-derived cholangiocytes.