title: Highly efficient generation of definitive endoderm lineage from human induced pluripotent stem cells.
abstract: BACKGROUND
  Although hepatocytes can be an option for liver transplantation, the shortage of donor organs continues to worsen. Since the development of induced pluripotent stem (iPS) cell technology, it is eagerly anticipated to produce functional elements from pluripotent stem cells. These functional cells differentiated from iPS cells could be used for transplantation, drug screening, and in vitro toxicology.
  
  
  METHODS
  Human iPS cells are maintained on Mitomycin C-treated mouse embryonic fibroblast layers in DMEM-Ham F12-based medium supplemented with Knockout Serum Replacement, nonessential amino acids, 2-mercaptoethanol, and Glutamax. Differentiation of human iPS cells into a definitive endodermal lineage was induced with PRMI 1640 medium supplemented with B27 and 100 ng/mL human activin A. Two B27 supplements were examined with and without insulin. Furthermore, the PI3 kinase inhibitor LY294002 was used to examine the effect of inhibiting insulin signaling.
  
  
  CONCLUSIONS
  We established efficient induction of definitive endodermal differentiation from iPS cells. Quantitative analysis revealed efficient (93.03 ± 2.74%) differentiation of human iPS cells into definitive endoderm cells using B27 minus insulin. This protocol may contribute as a fundamental technique to promote human iPS studies to develop cellular sources for transplantation.
authors.affiliation: Department of Regenerative Medicine, Graduate School of Medicine, Yokohama City University, Kanagawa, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Sekine
authors.forename: T
authors.initials: T
authors.lastname: Takebe
authors.forename: Y
authors.initials: Y
authors.lastname: Suzuki
authors.forename: A
authors.initials: A
authors.lastname: Kamiya
authors.forename: H
authors.initials: H
authors.lastname: Nakauchi
authors.forename: H
authors.initials: H
authors.lastname: Taniguchi
mesh_terms: D028341:Activins / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects; D002478:Cells, Cultured; D018920:Coculture Techniques; D003470:Culture Media / Q000737:chemistry / Q000378:metabolism; D004707:Endoderm / Q000166:cytology* / Q000378:metabolism / Q000637:transplantation; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000187:drug effects / Q000378:metabolism / Q000502:physiology* / Q000637:transplantation; D007328:Insulin / Q000378:metabolism; D051379:Mice; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D015398:Signal Transduction
chemical_list: D003470:Culture Media; D007328:Insulin; D000081082:Phosphoinositide-3 Kinase Inhibitors; D047428:Protein Kinase Inhibitors; C422405:activin A; D028341:Activins; D058539:Phosphatidylinositol 3-Kinase