Hepatocyte-like cells derived from human embryonic stem cells specifically via definitive endoderm and a progenitor stage.
Human embryonic stem cells offer a potential unlimited supply for functional hepatocytes, since they can differentiate into hepatocyte-like cells displaying a characteristic hepatic morphology and expressing various hepatic markers. These cells could be used in various applications such as studies of drug metabolism and hepatotoxicity, which however, would require a significant expression of drug metabolizing enzymes. To derive these cells we use a stepwise differentiation protocol where growth- and maturation factors are added. The first phase involves the formation of definitive endoderm. Next, these cells are treated with factors known to promote the induction and proliferation towards hepatic progenitor cell types. In the last phase the cells are terminally differentiated and maturated into functional hepatocyte-like cells. The cultures were characterized by analysis of endodermal or hepatic markers and compared to cultures derived without induction via definitive endoderm. Hepatic functions such as urea secretion, glycogen storage, indocyanine green uptake and secretion, and cytochrome P450-expression and activity were evaluated. The DE-Hep showed a hepatocyte morphology with sub-organized cells and exhibited many liver-functions including transporter activity and capacity to metabolize drugs specific for important cytochrome P450 sub-families. This represents an important step in differentiation of hESC into functional hepatocytes.
Brolén
Gabriella
G
Cellartis AB, Arvid Wallgrens Backe 20, 41346 Göteborg, Sweden.
Sivertsson
Louise
L
Björquist
Petter
P
Eriksson
Gustav
G
Ek
Monica
M
Semb
Henrik
H
Johansson
Inger
I
Andersson
Tommy B
TB
Ingelman-Sundberg
Magnus
M
Heins
Nico
N
D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002469:Cell Separation; D048430:Cell Shape / Q000187:drug effects; D004008:Diclofenac / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D004707:Endoderm / Q000166:cytology* / Q000187:drug effects; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D022781:Hepatocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008874:Midazolam / Q000378:metabolism; D010615:Phenacetin / Q000378:metabolism
D055415:Bone Morphogenetic Protein 4; D016222:Fibroblast Growth Factor 2; D004008:Diclofenac; D010615:Phenacetin; D008874:Midazolam