Engraftment potential of spheroid-forming hepatic endoderm derived from human embryonic stem cells.
Transplantation and drug discovery programs for liver diseases are hampered by the shortage of donor tissue. While recent studies have shown that hepatic cells can be derived from human embryonic stem cells (hESCs), few cases have shown selective enrichment of hESC-derived hepatocytes and their integration into host liver tissues. Here we demonstrate that the dissociation and reaggregation procedure after an endodermal differentiation of hESC produces spheroids mainly consisted of cells showing hepatic phenotypes in vitro and in vivo. A combined treatment with Wnt3a and bone morphogenic protein 4 efficiently differentiated hESCs into definitive endoderm in an adherent culture. Dissociation followed by reaggregation of these cells in a nonadherent condition lead to the isolation of spheroid-forming cells that preferentially expressed early hepatic markers from the adherent cell population. Further differentiation of these spheroid cells in the presence of the hepatocyte growth factor, oncostatin M, and dexamethasone produced a highly enriched population of cells exhibiting characteristics of early hepatocytes, including glycogen storage, indocyanine green uptake, and synthesis of urea and albumin. Furthermore, we show that grafted spheroid cells express hepatic features and attenuate the serum aspartate aminotransferase level in a model of acute liver injury. These data suggest that hepatic progenitor cells can be enriched by the spheroid formation of differentiating hESCs and that these cells have engraftment potential to replace damaged liver tissues.
Division of Biotechnology, Laboratory of Stem Cell Biology, College of Life Sciences and Biotechnology, Korea University, Seoul, Korea.
Sung-Eun
SE
Kim
Su Yeon
SY
An
Dong-Hun
DH
Woo
Jiyou
J
Han
Jong Hyun
JH
Kim
Yu Jin
YJ
Jang
Jeong Sang
JS
Son
Hyunwon
H
Yang
Yong Pil
YP
Cheon
Jong-Hoon
JH
Kim
D000418:Albumins / Q000096:biosynthesis; D000818:Animals; D015415:Biomarkers; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology; D002251:Carbon Tetrachloride; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D056486:Chemical and Drug Induced Liver Injury / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D003907:Dexamethasone / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D004707:Endoderm / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D006003:Glycogen / Q000096:biosynthesis; D006085:Graft Survival; D017228:Hepatocyte Growth Factor / Q000494:pharmacology; D022781:Hepatocytes / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D006801:Humans; D051379:Mice; D008819:Mice, Nude; D053683:Oncostatin M / Q000494:pharmacology; D018874:Spheroids, Cellular / Q000166:cytology / Q000378:metabolism / Q000637:transplantation*; D014183:Transplantation, Heterologous; D014508:Urea / Q000378:metabolism; D060509:Wnt3A Protein / Q000494:pharmacology
D000418:Albumins; C525809:BMP4 protein, human; D015415:Biomarkers; D055415:Bone Morphogenetic Protein 4; C066855:HGF protein, human; C555510:WNT3A protein, human; D060509:Wnt3A Protein; D053683:Oncostatin M; D017228:Hepatocyte Growth Factor; D003907:Dexamethasone; D014508:Urea; D006003:Glycogen; D002251:Carbon Tetrachloride