Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2
pubmed:dateCreated
2008-8-8
pubmed:abstractText
Hepatocytes derived from human embryonic stem cells (hESCs) are a potential cell source for regenerative medicine. However, the definitive factors that are responsible for hepatic differentiation of hESCs remain unclear. We aimed to evaluate the effects of various extracellular matrixes and growth factors on endodermal differentiation and to optimize the culture conditions to induce hepatic differentiation of hESCs. The transgene vector that contained enhanced green fluorescent protein (EGFP) under the control of human alpha-fetoprotein (AFP) enhancer/promoter was transfected into hESC lines. The transgenic hESCs were cultured on extracellular matrixes (collagen type I, laminin, and Matrigel) in the presence or absence of growth factors including hepatocyte growth factor (HGF), bone morphogenetic protein 4, fibroblast growth factor 4, all-trans-retinoic acid, and activin A. The expression of AFP-EGFP was measured by flow cytometry. The culture on Matrigel-coated dishes with 100 ng/ml activin A showed 19.5% of EGFP-positive proportions. Moreover, the sequential addition of 100 ng/ml activin A and 20 ng/ml HGF resulted in 21.7% and produced a higher yield of EGFP-positive cells than the group stimulated by activin A alone. RT-PCR and immunocytochemical staining revealed these EGFP-positive cells to differentiate into mesendoderm-like cells by use of activin A and then into hepatic endoderm cells by use of HGF. Two other hESC lines also differentiated into endoderm on the hepatic lineage by our method. In conclusion, we therefore found this protocol to effectively differentiate multiple hESC lines to early hepatocytes using activin A and HGF on Matrigel.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
http://linkedlifedata.com/resource/pubmed/chemical/Activins, http://linkedlifedata.com/resource/pubmed/chemical/BMP4 protein, human, http://linkedlifedata.com/resource/pubmed/chemical/Bone Morphogenetic Protein 4, http://linkedlifedata.com/resource/pubmed/chemical/Bone Morphogenetic Proteins, http://linkedlifedata.com/resource/pubmed/chemical/FGF4 protein, human, http://linkedlifedata.com/resource/pubmed/chemical/Fibroblast Growth Factor 4, http://linkedlifedata.com/resource/pubmed/chemical/Green Fluorescent Proteins, http://linkedlifedata.com/resource/pubmed/chemical/HGF protein, human, http://linkedlifedata.com/resource/pubmed/chemical/Hepatocyte Growth Factor, http://linkedlifedata.com/resource/pubmed/chemical/Tretinoin, http://linkedlifedata.com/resource/pubmed/chemical/activin A, http://linkedlifedata.com/resource/pubmed/chemical/alpha-Fetoproteins, http://linkedlifedata.com/resource/pubmed/chemical/enhanced green fluorescent protein
pubmed:status
MEDLINE
pubmed:month
Aug
pubmed:issn
0193-1857
pubmed:author
pubmed:issnType
Print
pubmed:volume
295
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
G313-21
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:18535293-Activins, pubmed-meshheading:18535293-Bone Morphogenetic Protein 4, pubmed-meshheading:18535293-Bone Morphogenetic Proteins, pubmed-meshheading:18535293-Carcinoma, Hepatocellular, pubmed-meshheading:18535293-Cell Differentiation, pubmed-meshheading:18535293-Embryonic Stem Cells, pubmed-meshheading:18535293-Endoderm, pubmed-meshheading:18535293-Extracellular Matrix, pubmed-meshheading:18535293-Fibroblast Growth Factor 4, pubmed-meshheading:18535293-Genetic Vectors, pubmed-meshheading:18535293-Green Fluorescent Proteins, pubmed-meshheading:18535293-Hepatocyte Growth Factor, pubmed-meshheading:18535293-Humans, pubmed-meshheading:18535293-Promoter Regions, Genetic, pubmed-meshheading:18535293-Reverse Transcriptase Polymerase Chain Reaction, pubmed-meshheading:18535293-Transgenes, pubmed-meshheading:18535293-Tretinoin, pubmed-meshheading:18535293-alpha-Fetoproteins
pubmed:year
2008
pubmed:articleTitle
Effects of extracellular matrixes and growth factors on the hepatic differentiation of human embryonic stem cells.
pubmed:affiliation
Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University, Japan. taishii@kuhp.kyoto-u.ac.jp
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't