New markers for tracking endoderm induction and hepatocyte differentiation from human pluripotent stem cells.
The efficient generation of hepatocytes from human pluripotent stem cells (hPSCs) requires the induction of a proper endoderm population, broadly characterized by the expression of the cell surface marker CXCR4. Strategies to identify and isolate endoderm subpopulations predisposed to the liver fate do not exist. In this study, we generated mouse monoclonal antibodies against human embryonic stem cell-derived definitive endoderm with the goal of identifying cell surface markers that can be used to track the development of this germ layer and its specification to a hepatic fate. Through this approach, we identified two endoderm-specific antibodies, HDE1 and HDE2, which stain different stages of endoderm development and distinct derivative cell types. HDE1 marks a definitive endoderm population with high hepatic potential, whereas staining of HDE2 tracks with developing hepatocyte progenitors and hepatocytes. When used in combination, the staining patterns of these antibodies enable one to optimize endoderm induction and hepatic specification from any hPSC line.
McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada M5G 1L7.
Audrey
A
Holtzinger
Oregon Stem Cell Center, Oregon Health & Science University, Portland, OR 97239-3098, USA.
Philip R
PR
Streeter
McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada M5G 1L7.
Farida
F
Sarangi
McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada M5G 1L7.
Scott
S
Hillborn
McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada M5G 1L7.
Maryam
M
Niapour
McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada M5G 1L7.
Shinichiro
S
Ogawa
McEwen Centre for Regenerative Medicine, University Health Network, Toronto, Ontario, Canada M5G 1L7 Department of Medical Biophysics, University of Toronto, Toronto, Ontario, Canada M5G 2M9 Princess Margaret Cancer Centre, Toronto, Ontario, Canada M5T 2M9 gkeller@uhnresearch.ca.
Gordon
G
Keller
D000818:Animals; D000906:Antibodies / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism*; D002454:Cell Differentiation; D002460:Cell Line; D002469:Cell Separation; D004475:Ectoderm / Q000166:cytology; D004707:Endoderm / Q000166:cytology*; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007700:Kinetics; D008648:Mesoderm / Q000166:cytology; D008807:Mice, Inbred BALB C; D010179:Pancreas / Q000196:embryology; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D013194:Staining and Labeling
D000906:Antibodies; D015415:Biomarkers
Endoderm marker; Hepatocyte differentiation; Human pluripotent stem cell