title: Factors from human embryonic stem cell-derived fibroblast-like cells promote topology-dependent hepatic differentiation in primate embryonic and induced pluripotent stem cells.
abstract: The future clinical use of embryonic stem cell (ESC)-based hepatocyte replacement therapy depends on the development of an efficient procedure for differentiation of hepatocytes from ESCs. Here we report that a high density of human ESC-derived fibroblast-like cells (hESdFs) supported the efficient generation of hepatocyte-like cells with functional and mature hepatic phenotypes from primate ESCs and human induced pluripotent stem cells. Molecular and immunocytochemistry analyses revealed that hESdFs caused a rapid loss of pluripotency and induced a sequential endoderm-to-hepatocyte differentiation in the central area of ESC colonies. Knockdown experiments demonstrated that pluripotent stem cells were directed toward endodermal and hepatic lineages by FGF2 and activin A secreted from hESdFs. Furthermore, we found that the central region of ESC colonies was essential for the hepatic endoderm-specific differentiation, because its removal caused a complete disruption of endodermal differentiation. In conclusion, we describe a novel in vitro differentiation model and show that hESdF-secreted factors act in concert with regional features of ESC colonies to induce robust hepatic endoderm differentiation in primate pluripotent stem cells.
authors: From the Divisions of Medical Research, Taipei 10002, Taiwan.
authors: Hsiang-Po
authors: HP
authors: Huang
authors: Reproductive Endocrinology and Infertility, Taipei 10002, Taiwan.
authors: Chun-Ying
authors: CY
authors: Yu
authors: Reproductive Endocrinology and Infertility, Taipei 10002, Taiwan; Institute of Clinical Genomics, Taipei 10617, Taiwan.
authors: Hsin-Fu
authors: HF
authors: Chen
authors: From the Divisions of Medical Research, Taipei 10002, Taiwan.
authors: Pin-Hsun
authors: PH
authors: Chen
authors: Genomics Research Center, Taipei 11574, Taiwan.
authors: Ching-Yu
authors: CY
authors: Chuang
authors: Departments of Dermatology, Taipei 10002, Taiwan; Biomedical Engineering, Taipei 10617, Taiwan.
authors: Sung-Jan
authors: SJ
authors: Lin
authors: Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 11574, Taiwan.
authors: Shih-Tsung
authors: ST
authors: Huang
authors: Anesthesiology, National Taiwan University Hospital, Taipei 10002, Taiwan.
authors: Wei-Hung
authors: WH
authors: Chan
authors: Institute of Toxicology, National Taiwan University, Taipei 10617, Taiwan.
authors: Tzuu-Huei
authors: TH
authors: Ueng
authors: Reproductive Endocrinology and Infertility, Taipei 10002, Taiwan; Institute of Clinical Genomics, Taipei 10617, Taiwan.
authors: Hong-Nerng
authors: HN
authors: Ho
authors: Genomics Research Center, Taipei 11574, Taiwan; Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 11574, Taiwan; Institute of Clinical Medicine, Taipei Medical University, Taipei 11031, Taiwan. Electronic address: kuohuch@gate.sinica.edu.tw.
authors: Hung-Chih
authors: HC
authors: Kuo
mesh_terms: D028341:Activins / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism*; D000882:Haplorhini; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D051379:Mice
chemical_list: C422405:activin A; D016222:Fibroblast Growth Factor 2; D028341:Activins