Factors from human embryonic stem cell-derived fibroblast-like cells promote topology-dependent hepatic differentiation in primate embryonic and induced pluripotent stem cells.
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.
Huang
Hsiang-Po
HP
From the Divisions of Medical Research, Taipei 10002, Taiwan.
Yu
Chun-Ying
CY
Reproductive Endocrinology and Infertility, Taipei 10002, Taiwan.
Chen
Hsin-Fu
HF
Reproductive Endocrinology and Infertility, Taipei 10002, Taiwan; Institute of Clinical Genomics, Taipei 10617, Taiwan.
Chen
Pin-Hsun
PH
From the Divisions of Medical Research, Taipei 10002, Taiwan.
Chuang
Ching-Yu
CY
Genomics Research Center, Taipei 11574, Taiwan.
Lin
Sung-Jan
SJ
Departments of Dermatology, Taipei 10002, Taiwan; Biomedical Engineering, Taipei 10617, Taiwan.
Huang
Shih-Tsung
ST
Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 11574, Taiwan.
Chan
Wei-Hung
WH
Anesthesiology, National Taiwan University Hospital, Taipei 10002, Taiwan.
Ueng
Tzuu-Huei
TH
Institute of Toxicology, National Taiwan University, Taipei 10617, Taiwan.
Ho
Hong-Nerng
HN
Reproductive Endocrinology and Infertility, Taipei 10002, Taiwan; Institute of Clinical Genomics, Taipei 10617, Taiwan.
Kuo
Hung-Chih
HC
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.
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
C422405:activin A; D016222:Fibroblast Growth Factor 2; D028341:Activins