Generation of functional human hepatic endoderm from human induced pluripotent stem cells.
With the advent of induced pluripotent stem cell (iPSC) technology, it is now feasible to generate iPSCs with a defined genotype or disease state. When coupled with direct differentiation to a defined lineage, such as hepatic endoderm (HE), iPSCs would revolutionize the way we study human liver biology and generate efficient "off the shelf" models of human liver disease. Here, we show the "proof of concept" that iPSC lines representing both male and female sexes and two ethnic origins can be differentiated to HE at efficiencies of between 70%-90%, using a method mimicking physiological relevant condition. The iPSC-derived HE exhibited hepatic morphology and expressed the hepatic markers albumin and E-cadherin, as assessed by immunohistochemistry. They also expressed alpha-fetoprotein, hepatocyte nuclear factor-4a, and a metabolic marker, cytochrome P450 7A1 (Cyp7A1), demonstrating a definitive endodermal lineage differentiation. Furthermore, iPSC-derived hepatocytes produced and secreted the plasma proteins, fibrinogen, fibronectin, transthyretin, and alpha-fetoprotein, an essential feature for functional HE. Additionally iPSC-derived HE supported both CYP1A2 and CYP3A4 metabolism, which is essential for drug and toxicology testing.


CONCLUSIONS
This work is first to demonstrate the efficient generation of hepatic endodermal lineage from human iPSCs that exhibits key attributes of hepatocytes, and the potential application of iPSC-derived HE in studying human liver biology. In particular, iPSCs from individuals representing highly polymorphic variants in metabolic genes and different ethnic groups will provide pharmaceutical development and toxicology studies a unique opportunity to revolutionize predictive drug toxicology assays and allow the creation of in vitro hepatic disease models.
Sullivan
Gareth J
GJ
Medical Research Council (MRC) Centre for Regenerative Medicine, University of Edinburgh, Edinburgh, UK. gsulliva@staffmail.ed.ac.uk
Hay
David C
DC
Park
In-Hyun
IH
Fletcher
Judy
J
Hannoun
Zara
Z
Payne
Catherine M
CM
Dalgetty
Donna
D
Black
James R
JR
Ross
James A
JA
Samuel
Kay
K
Wang
Gang
G
Daley
George Q
GQ
Lee
Je-Hyuk
JH
Church
George M
GM
Forbes
Stuart J
SJ
Iredale
John P
JP
Wilmut
Ian
I
D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology*; D019070:Cell Lineage; D004707:Endoderm / Q000166:cytology*; D005260:Female; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D008099:Liver / Q000166:cytology*; D008297:Male