Small-molecule-driven hepatocyte differentiation of human pluripotent stem cells.
The differentiation of pluripotent stem cells to hepatocytes is well established, yet current methods suffer from several drawbacks. These include a lack of definition and reproducibility, which in part stems from continued reliance on recombinant growth factors. This has remained a stumbling block for the translation of the technology into industry and the clinic for reasons associated with cost and quality. We have devised a growth-factor-free protocol that relies on small molecules to differentiate human pluripotent stem cells toward a hepatic phenotype. The procedure can efficiently direct both human embryonic stem cells and induced pluripotent stem cells to hepatocyte-like cells. The final population of cells demonstrates marker expression at the transcriptional and protein levels, as well as key hepatic functions such as serum protein production, glycogen storage, and cytochrome P450 activity.
Department of Biochemistry, Faculty of Medicine, Institute of Basic Medical Sciences, University of Oslo, PO Box 1112 Blindern, 0317 Oslo, Norway.
Richard
R
Siller
Department of Biochemistry, Faculty of Medicine, Institute of Basic Medical Sciences, University of Oslo, PO Box 1112 Blindern, 0317 Oslo, Norway.
Sebastian
S
Greenhough
Department of Biochemistry, Faculty of Medicine, Institute of Basic Medical Sciences, University of Oslo, PO Box 1112 Blindern, 0317 Oslo, Norway.
Elena
E
Naumovska
Department of Biochemistry, Faculty of Medicine, Institute of Basic Medical Sciences, University of Oslo, PO Box 1112 Blindern, 0317 Oslo, Norway; Norwegian Center for Stem Cell Research, PO Box 1112 Blindern, 0317 Oslo, Norway; Institute of Immunology, Oslo University Hospital-Rikshospitalet, PO Box 4950 Nydalen, Oslo 0424, Norway. Electronic address: gareth.sullivan@medisin.uio.no.
Gareth J
GJ
Sullivan
D001798:Blood Proteins / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D003907:Dexamethasone / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D006003:Glycogen / Q000378:metabolism; D038362:Glycogen Synthase Kinase 3 / Q000037:antagonists & inhibitors / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D006801:Humans; D008856:Microscopy, Fluorescence; D009842:Oligopeptides / Q000494:pharmacology; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D019859:Proto-Oncogene Proteins c-met / Q000819:agonists / Q000378:metabolism; D054852:Small Molecule Libraries / Q000494:pharmacology*
D001798:Blood Proteins; D009842:Oligopeptides; D054852:Small Molecule Libraries; D003907:Dexamethasone; D006003:Glycogen; C578458:N-hexanoic-Tyr-Ile-(6) aminohexanoic amide; D019859:Proto-Oncogene Proteins c-met; D038362:Glycogen Synthase Kinase 3; D004121:Dimethyl Sulfoxide