title: Highly efficient differentiation of functional hepatocytes from human induced pluripotent stem cells.
abstract: Human induced pluripotent stem cells (hiPSCs) hold great potential for use in regenerative medicine, novel drug development, and disease progression/developmental studies. Here, we report highly efficient differentiation of hiPSCs toward a relatively homogeneous population of functional hepatocytes. hiPSC-derived hepatocytes (hiHs) not only showed a high expression of hepatocyte-specific proteins and liver-specific functions, but they also developed a functional biotransformation system including phase I and II metabolizing enzymes and phase III transporters. Nuclear receptors, which are critical for regulating the expression of metabolizing enzymes, were also expressed in hiHs. hiHs also responded to different compounds/inducers of cytochrome P450 as mature hepatocytes do. To follow up on this observation, we analyzed the drug metabolizing capacity of hiHs in real time using a novel ultra performance liquid chromatography-tandem mass spectrometry. We found that, like freshly isolated primary human hepatocytes, the seven major metabolic pathways of the drug bufuralol were found in hiHs. In addition, transplanted hiHs engrafted, integrated, and proliferated in livers of an immune-deficient mouse model, and secreted human albumin, indicating that hiHs also function in vivo. In conclusion, we have generated a method for the efficient generation of hepatocytes from induced pluripotent stem cells in vitro and in vivo, and it appears that the cells function similarly to primary human hepatocytes, including developing a complete metabolic function. These results represent a significant step toward using patient/disease-specific hepatocytes for cell-based therapeutics as well as for pharmacology and toxicology studies.
authors.affiliation: Department of Internal Medicine, University of California, Sacramento, CA, USA.
authors.forename: Xiaocui
authors.initials: X
authors.lastname: Ma
authors.forename: Yuyou
authors.initials: Y
authors.lastname: Duan
authors.forename: Benjamin
authors.initials: B
authors.lastname: Tschudy-Seney
authors.forename: Garrett
authors.initials: G
authors.lastname: Roll
authors.forename: Iman Saramipoor
authors.initials: IS
authors.lastname: Behbahan
authors.forename: Tijess P
authors.initials: TP
authors.lastname: Ahuja
authors.forename: Vladimir
authors.initials: V
authors.lastname: Tolstikov
authors.forename: Charles
authors.initials: C
authors.lastname: Wang
authors.forename: Jeannine
authors.initials: J
authors.lastname: McGee
authors.forename: Shiva
authors.initials: S
authors.lastname: Khoobyari
authors.forename: Jan A
authors.initials: JA
authors.lastname: Nolta
authors.forename: Holger
authors.initials: H
authors.lastname: Willenbring
authors.forename: Mark A
authors.initials: MA
authors.lastname: Zern
mesh_terms: D000319:Adrenergic beta-Antagonists / Q000378:metabolism; D000418:Albumins / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D004983:Ethanolamines / Q000378:metabolism; D015870:Gene Expression; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation*; D006801:Humans; D016867:Immunocompromised Host; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008099:Liver; D050216:Metabolic Detoxication, Phase I / Q000502:physiology*; D050217:Metabolic Detoxication, Phase II / Q000502:physiology*; D051379:Mice; D016513:Mice, SCID; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D014183:Transplantation, Heterologous
chemical_list: D000319:Adrenergic beta-Antagonists; D000418:Albumins; D015415:Biomarkers; D004983:Ethanolamines; D018160:Receptors, Cytoplasmic and Nuclear; C010831:bufuralol; D003577:Cytochrome P-450 Enzyme System
keywords: Hepatocyte differentiation; Induced pluripotent stem cells; Liver regeneration; Stem cell transplantation