TY - JOUR
T1 - Instant hepatic differentiation of human embryonic stem cells using activin A and a deleted variant of HGF
AU - Chen, Yong
AU - Soto-Gutierrez, Alejandro
AU - Navarro-Alvarez, Nalu
AU - Rivas-Carrillo, Jorge David
AU - Yamatsuji, Tomoki
AU - Shirakawa, Yasuhiro
AU - Tanaka, Noriaki
AU - Kobayashi, Naoya
PY - 2006
Y1 - 2006
N2 - 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.
AB - 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.
KW - Differentiation
KW - Hepatocyte
KW - Hepatocyte growth factor
KW - hES cells
UR - http://www.scopus.com/inward/record.url?scp=33846600941&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33846600941&partnerID=8YFLogxK
U2 - 10.3727/000000006783981305
DO - 10.3727/000000006783981305
M3 - Article
C2 - 17299990
AN - SCOPUS:33846600941
SN - 0963-6897
VL - 15
SP - 865
EP - 871
JO - Cell Transplantation
JF - Cell Transplantation
IS - 10
ER -