Differentiation of human embryonic stem cells into hepatocytes in 2D and 3D culture systems in vitro.
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The differentiation of hESCs into hepatocyte-like cells within 3D collagen scaffolds containing exogenous growth factors gives rise to cells displaying morphological features, gene expression patterns and metabolic activities characteristic of hepatocytes and may provide a source of differentiated cells for treatment of liver diseases.Abstract:
Human embryonic stem cells (hESCs) have enormous potential as a source of cells for cell replacement therapies and as a model for early human development. In this study we examined the differentiating potential of hESCs into hepatocytes in two- and three-dimensional (2D and 3D) culture systems. Embryoid bodies (EBs) were inserted into a collagen scaffold 3D culture system or cultured on collagen-coated dishes and stimulated with exogenous growth factors to induce hepatic histogenesis. Immunofluorescence analysis revealed the expression of albumin (ALB) and cytokeratin-18 (CK-18). The differentiated cells in 2D and 3D culture system displayed several characteristics of hepatocytes, including expression of transthyretin, alpha-1-antitrypsin, cytokeratin 8, 18, 19, tryptophan-2,3-dioxygenase, tyrosine aminotransferase, glucose-6-phosphatase (G6P), cytochrome P450 subunits 7a1 and secretion of alpha-fetoprotein (AFP) and ALB and production of urea. In 3D culture, ALB and G6P were detected earlier and higher levels of urea and AFP were produced, when compared with 2D culture. Electron microscopy of differentiated hESCs showed hepatocyte-like ultrastructure, including glycogon granules, well-developed Golgi apparatuses, rough and smooth endoplasmic reticuli and intercellular canaliculi. The differentiation of hESCs into hepatocyte-like cells within 3D collagen scaffolds containing exogenous growth factors, gives rise to cells displaying morphological features, gene expression patterns and metabolic activities characteristic of hepatocytes and may provide a source of differentiated cells for treatment of liver diseases.read more
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Three-Dimensional Cell Culture Systems and Their Applications in Drug Discovery and Cell-Based Biosensors
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Joseph Itskovitz-Eldor,Maya Schuldiner,Dorit Karsenti,Amir Eden,Ofra Yanuka,Michal Amit,Hermona Soreq,Nissim Benvenisty +7 more
TL;DR: The ability to induce formation of human embryoid bodies that contain cells of neuronal, hematopoietic and cardiac origins will be useful in studying early human embryonic development as well as in transplantation medicine.
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Effects of eight growth factors on the differentiation of cells derived from human embryonic stem cells
TL;DR: This analysis sets the stage for directing differentiation of human ES cells in culture and indicates that multiple human cell types may be enriched in vitro by specific factors.
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Treatment of the Crigler–Najjar Syndrome Type I with Hepatocyte Transplantation
Ira J. Fox,Jayanta Roy Chowdhury,Stuart S. Kaufman,Timothy C. Goertzen,Namita Roy Chowdhury,Phyllis I. Warkentin,Kenneth Dorko,Bernhard V. Sauter,Stephen C. Strom +8 more
TL;DR: Although phototherapy successfully reduces serum bilirubin levels, patients are again at risk for kernicterus around the time of puberty, when phototherapy becomes less effective, and liver transplantation is the only definitive treatment.
Journal ArticleDOI
Differentiation of human embryonic stem cells on three-dimensional polymer scaffolds.
Shulamit Levenberg,Ngan F. Huang,Erin B. Lavik,Arlin B. Rogers,Joseph Itskovitz-Eldor,Robert Langer +5 more
TL;DR: It is shown that complex structures with features of various committed embryonic tissues can be generated, in vitro, by using early differentiating hES cells and further inducing their differentiation in a supportive 3D environment such as poly(lactic-co-glycolic acid)/poly(l-lactic acid) polymer scaffolds.
Journal ArticleDOI
Initiation of Mammalian Liver Development from Endoderm by Fibroblast Growth Factors
TL;DR: Different FGF signals appear to initiate distinct phases of liver development during mammalian organogenesis, and studies with FGFs and their specific inhibitors showed that FGF8 contributes to the morphogenetic outgrowth of the hepatic endoderm.