Instant hepatic differentiation of human embryonic stem cells using activin A and a deleted variant of HGF.
Human embryonic stem (hES) cells have the ability to differentiate into a variety of different cell lineages and potentially provide a source of differentiated cells for many therapeutic uses. Here we investigated an efficient method of hepatic differentiation from hES cells. A human ES cell line, KhES-1, was used and maintained by a nonfeeder method. KhES-1 cells were cultured for 5 days in the presence of human activin A (50 ng/ml) and then treated with a deleted variant of hepatocyte growth factor (dHGF) at 0, 100, or 500 ng/ml for 7 days. The resultant cells were biologically analyzed. The expression of the endodermal genes SOX17 and FOXA2 increased in KhES-1 cells after activin A treatment. In contrast, Oct4, a self-renewal undifferentiated marker, decreased in a time-dependent manner in KhES-1 cells. Following a 7-day treatment of the resultant cells with dHGF, especially at 500 ng/ml, KhES-1 cells showed an expression of the hepatic makers albumin, AFP, and CK18. Transitional electron microscopy showed well-developed glycogen rosettes and a gap junction in KhES-1 cells treated with 500 ng/ml of dHGF. We developed an efficient method to differentiate KhES-1 cells into hepatocyte-like cells in vitro using 50 ng/ml of activin A and 500 ng/ml of dHGF.
Chen
Yong
Y
Department of Surgery, Okayama University Graduate School of Medicine and Dentistry, Okayama 700-8558, Japan.
Soto-Gutierrez
Alejandro
A
Navarro-Alvarez
Nalu
N
Rivas-Carrillo
Jorge David
JD
Yamatsuji
Tomoki
T
Shirakawa
Yasuhiro
Y
Tanaka
Noriaki
N
Basma
Hesham
H
Fox
Ira J
IJ
Kobayashi
Naoya
N
D028341:Activins / Q000494:pharmacology*; D000418:Albumins / Q000378:metabolism; D000641:Ammonia / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000648:ultrastructure; D006003:Glycogen / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000235:genetics / Q000494:pharmacology*; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D006801:Humans; D008012:Lidocaine / Q000378:metabolism; D046529:Microscopy, Electron, Transmission; D009154:Mutation
D000418:Albumins; D028341:Activins; D017228:Hepatocyte Growth Factor; D000641:Ammonia; D006003:Glycogen; D008012:Lidocaine