title: Hepatic differentiation of human pluripotent stem cells in miniaturized format suitable for high-throughput screen.
abstract: The establishment of protocols to differentiate human pluripotent stem cells (hPSCs) including embryonic (ESC) and induced pluripotent (iPSC) stem cells into functional hepatocyte-like cells (HLCs) creates new opportunities to study liver metabolism, genetic diseases and infection of hepatotropic viruses (hepatitis B and C viruses) in the context of specific genetic background. While supporting efficient differentiation to HLCs, the published protocols are limited in terms of differentiation into fully mature hepatocytes and in a smaller-well format. This limitation handicaps the application of these cells to high-throughput assays. Here we describe a protocol allowing efficient and consistent hepatic differentiation of hPSCs in 384-well plates into functional hepatocyte-like cells, which remain differentiated for more than 3weeks. This protocol affords the unique opportunity to miniaturize the hPSC-based differentiation technology and facilitates screening for molecules in modulating liver differentiation, metabolism, genetic network, and response to infection or other external stimuli.
authors.affiliation: Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health (NIH), Bethesda, MD 20892, USA. Electronic address: arnaud.carpentier@twincore.de.
authors.forename: Arnaud
authors.initials: A
authors.lastname: Carpentier
authors.affiliation: Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health (NIH), Bethesda, MD 20892, USA.
authors.forename: Ila
authors.initials: I
authors.lastname: Nimgaonkar
authors.affiliation: Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health (NIH), Bethesda, MD 20892, USA.
authors.forename: Virginia
authors.initials: V
authors.lastname: Chu
authors.affiliation: Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health (NIH), Bethesda, MD 20892, USA.
authors.forename: Yuchen
authors.initials: Y
authors.lastname: Xia
authors.affiliation: Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health (NIH), Bethesda, MD 20892, USA.
authors.forename: Zongyi
authors.initials: Z
authors.lastname: Hu
authors.affiliation: Liver Diseases Branch, National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), National Institutes of Health (NIH), Bethesda, MD 20892, USA. Electronic address: jakel@bdg10.niddk.nih.gov.
authors.forename: T Jake
authors.initials: TJ
authors.lastname: Liang
mesh_terms: D002454:Cell Differentiation; D051544:Cytochrome P-450 CYP3A / Q000378:metabolism; D051557:Hepatocyte Nuclear Factor 4 / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence; D008904:Miniaturization; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D000509:alpha-Fetoproteins / Q000378:metabolism
chemical_list: C496592:HNF4A protein, human; D051557:Hepatocyte Nuclear Factor 4; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D000509:alpha-Fetoproteins; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human
keywords: 384-Well plates; Hepatic differentiation; High-throughput assay; Pluripotent stem cells