Generation of metabolically functioning hepatocytes from human pluripotent stem cells by FOXA2 and HNF1α transduction.
OBJECTIVE
Hepatocyte-like cells differentiated from human embryonic stem cells (hESCs) and induced pluripotent stem cells (hiPSCs) can be utilized as a tool for screening for hepatotoxicity in the early phase of pharmaceutical development. We have recently reported that hepatic differentiation is promoted by sequential transduction of SOX17, HEX, and HNF4α into hESC- or hiPSC-derived cells, but further maturation of hepatocyte-like cells is required for widespread use of drug screening.


METHODS
To screen for hepatic differentiation-promoting factors, we tested the seven candidate genes related to liver development.


RESULTS
The combination of two transcription factors, FOXA2 and HNF1α, promoted efficient hepatic differentiation from hESCs and hiPSCs. The expression profile of hepatocyte-related genes (such as genes encoding cytochrome P450 enzymes, conjugating enzymes, hepatic transporters, and hepatic nuclear receptors) achieved with FOXA2 and HNF1α transduction was comparable to that obtained in primary human hepatocytes. The hepatocyte-like cells generated by FOXA2 and HNF1α transduction exerted various hepatocyte functions including albumin and urea secretion, and the uptake of indocyanine green and low density lipoprotein. Moreover, these cells had the capacity to metabolize all nine tested drugs and were successfully employed to evaluate drug-induced cytotoxicity.


CONCLUSIONS
Our method employing the transduction of FOXA2 and HNF1α represents a useful tool for the efficient generation of metabolically functional hepatocytes from hESCs and hiPSCs, and the screening of drug-induced cytotoxicity.
Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Osaka 565-0871, Japan.
Kazuo
K
Takayama
Mitsuru
M
Inamura
Kenji
K
Kawabata
Michiko
M
Sugawara
Kiyomi
K
Kikuchi
Maiko
M
Higuchi
Yasuhito
Y
Nagamoto
Hitoshi
H
Watanabe
Katsuhisa
K
Tashiro
Fuminori
F
Sakurai
Takao
T
Hayakawa
Miho Kusuda
MK
Furue
Hiroyuki
H
Mizuguchi
D016642:Bupropion / Q000378:metabolism; D002454:Cell Differentiation; D002460:Cell Line; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D004983:Ethanolamines / Q000378:metabolism; D018014:Gene Transfer Techniques; D051538:Hepatocyte Nuclear Factor 1-alpha / Q000235:genetics* / Q000378:metabolism; D051542:Hepatocyte Nuclear Factor 3-beta / Q000235:genetics* / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D008874:Midazolam / Q000378:metabolism; D017239:Paclitaxel / Q000378:metabolism; D010615:Phenacetin / Q000378:metabolism; D013739:Testosterone / Q000378:metabolism; D014044:Tolbutamide / Q000378:metabolism; D014161:Transduction, Genetic
D004983:Ethanolamines; C496502:FOXA2 protein, human; C496487:HNF1A protein, human; D051538:Hepatocyte Nuclear Factor 1-alpha; D016642:Bupropion; D051542:Hepatocyte Nuclear Factor 3-beta; D013739:Testosterone; C010831:bufuralol; D014044:Tolbutamide; D010615:Phenacetin; D017239:Paclitaxel; D008874:Midazolam