Isolation of hepatocyte-like cells from mouse embryoid body cells.
We previously reported that embryoid body (EB) cells derived from embryonic stem (ES) cells are capable of differentiating into functional hepatocyte-like cells both in vitro and in vivo. Because transplantation of EB-derived cells into the liver via the spleen resulted in a low incidence of teratoma formation, purification of hepatocyte-like cells is required to prevent teratoma formation. The aim of this study was to purify hepatocyte-like cells from cultured EBs. For the isolation of hepatocyte-like cells, EBs cultured for 15 days were treated with trypsin-EDTA. The disaggregated cells were plated on a gelatin-coated dish as a monolayer. These cells were separated by Percoll gradient centrifugation, enriched by magnetic cell sorting, and purified by FACS. The purified hepatocyte-like cells in monolayer cultures were positive for immunostaining for albumin and expressed albumin mRNA, but not Oct3/4 mRNA. Transplantation of the purified hepatocyte-like cells derived from mouse ES cells might be an effective treatment for liver failure.
Kumashiro
Y
Y
Department of Hepato-Biliary-Pancreatic Surgery, Graduate School of Medicine and Dentistry, Tokyo Medical and Dental University, Tokyo, Japan.
Teramoto
K
K
Shimizu-Saito
K
K
Asahina
K
K
Teraoka
H
H
Arii
S
S
D000818:Animals; D001483:Base Sequence; D002454:Cell Differentiation; D002460:Cell Line; D002469:Cell Separation / Q000379:methods; D017931:DNA Primers; D005434:Flow Cytometry; D022781:Hepatocytes / Q000166:cytology*; D008099:Liver / Q000196:embryology*; D051379:Mice; D013234:Stem Cells / Q000166:cytology*
D017931:DNA Primers