title: Albumin gene targeting in human embryonic stem cells and induced pluripotent stem cells with helper-dependent adenoviral vector to monitor hepatic differentiation.
abstract: Although progresses in developing differentiation procedures have been achieved, it remains challenging to generate hES/iPS cell-derived mature hepatocytes. We performed knock-in of a monomeric Kusabira orange (mKO1) cassette in the albumin (ALB) gene, in human embryonic stem (hES) cells and induced pluripotent stem (hiPS) cells, with the use of the helper-dependent adenovirus vector (HDAdV). Upon induction into the hepatic lineages, these knock-in hES/iPS cells differentiated into cells that displayed several known hepatic functions. The mKO1 knock-in (ALB/mKo1) hES/hiPS cells were used to visualize hepatic differentiation in vitro. mKO1 reporter expression recapitulated endogenous ALB transcriptional activity. ALB/mKo1 [Hi] population isolated by flow cytometry was confirmed to be enriched with ALB mRNA. Expression profile analyses revealed that characteristic hepatocyte genes and genes related to drug metabolism and many aspects of liver function were highly enriched in the ALB/mKo1 [Hi] population. Our data demonstrate that ALB/mKo1 knock-in hES/iPS cells are valuable resources for monitoring in vitro hepatic differentiation, isolation and analyses of hES and hiPS cells-derived hepatic cells that actively transcribing ALB. These knock-in hES/iPS cell lines could provide further insights into the mechanism of hepatic differentiation and molecular signatures of the hepatic cells derived from hES/iPS cells.
authors.affiliation: Department of Stem Cell Biology, Institute of Molecular Embryology and Genetics, Kumamoto University, Honjo 2-2-1, Kumamoto 860-0811, Japan.
authors.forename: Kahoko
authors.initials: K
authors.lastname: Umeda
authors.forename: Keiichiro
authors.initials: K
authors.lastname: Suzuki
authors.forename: Taiji
authors.initials: T
authors.lastname: Yamazoe
authors.forename: Nobuaki
authors.initials: N
authors.lastname: Shiraki
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Higuchi
authors.forename: Kumiko
authors.initials: K
authors.lastname: Tokieda
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Kume
authors.forename: Kohnosuke
authors.initials: K
authors.lastname: Mitani
authors.forename: Shoen
authors.initials: S
authors.lastname: Kume
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000418:Albumins / Q000235:genetics*; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D019070:Cell Lineage / Q000235:genetics; D002469:Cell Separation; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D055879:Gene Knock-In Techniques; D018390:Gene Targeting; D056426:Genetic Loci / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D006378:Helper Viruses / Q000235:genetics; D006801:Humans; D008658:Inactivation, Metabolic / Q000235:genetics; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D008099:Liver / Q000166:cytology*; D008164:Luminescent Proteins / Q000378:metabolism; D008297:Male; D000097573:Red Fluorescent Protein
chemical_list: D000418:Albumins; D015415:Biomarkers; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein