Differentiation and enrichment of hepatocyte-like cells from human embryonic stem cells in vitro and in vivo.
Human embryonic stem cells (hESC) may provide a cell source for functional hepatocytes. The aim of this study is to establish a viable human hepatocyte-like cell line from hESC that can be used for cell-based therapies. The differentiated hESC were enriched by transducing with a lentivirus vector containing the green fluorescent protein (GFP) gene driven by the alpha1-antitrypsin promoter; the GFP gene is expressed in committed hepatocyte progenitors and hepatocytes. GFP+ hESC were purified by laser microdissection and pressure catapulting. In addition, differentiated hESC that were transduced with a lentivirus triple-fusion vector were transplanted into NOD-SCID mice, and the luciferase-induced bioluminescence in the livers was evaluated by a charge-coupled device camera. GFP+ hESC expressed a large series of liver-specific genes, and expression levels of these genes were significantly improved by purifying GFP+ hESC; our results demonstrated that purified differentiated hESC express nearly physiological levels of liver-specific genes and have liver-specific functions that are comparable to those of primary human hepatocytes. The differentiated hESC survived and engrafted in mouse livers, and human liver-specific mRNA and protein species were detected in the transplanted mouse liver and serum at 3 weeks after transplantation. This is the first time that human albumin generated by hESC-derived hepatocytes was detected in the serum of an animal model. This also represents the first successful transplantation of differentiated hESC in an animal liver and the first bioluminescence imaging of hESC in the liver. This study is an initial step in establishing a viable hepatocyte-like cell line from hESC. Disclosure of potential conflicts of interest is found at the end of this article.
Duan
Yuyou
Y
Transplant Research Institute, University of California Davis Medical Center, 4635 2nd Avenue, Suite 1001, Sacramento, California 95817, USA.
Catana
Andreea
A
Meng
Ying
Y
Yamamoto
Naoki
N
He
Songqing
S
Gupta
Sanjeev
S
Gambhir
Sanjiv Sam
SS
Zern
Mark A
MA
D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000473:pathology; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology / Q000502:physiology* / Q000637:transplantation*; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D006085:Graft Survival; D022781:Hepatocytes / Q000473:pathology / Q000502:physiology* / Q000637:transplantation*; D006801:Humans; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID