In Vitro Differentiation of Embryonic Stem Cells into Hepatocyte‐Like Cells Identified by Cellular Uptake of Indocyanine Green

Abstract

Background and Aims. Embryonic stem (ES) cells have a pluripotent ability to differentiate into a variety of cell lineages in vitro.We have recently found the emergence of cell clusters that show the cellular uptake of indocyanine green (ICG) in the culture of differentiated ES cells. ICG is clinically used as a test substance to evaluate liver function because it is eliminated exclusively by hepatocytes. The aim of the present study was to investigate the hepatic characteristics of ICG-stained cells.Methods. Embryoid bodies (EBs), formed by a 5-day hanging drop culture of ES cells, were allowed to outgrow in the placed culture. Gene expression of hepatocyte markers was analyzed by reverse transcriptase-polymerase chain reaction, and albumin production was examined immunohistochemically. Morphology and cellular components were investigated by electron microscopy. ICG-stained cells were further transplanted into the portal vein of mice.

Differentiation of mouse embryonic stem cells to hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines

“…that ES cells can be differentiated into hepatocyte-like cells [1][2][3][4][5][6] . ES cell-derived hepatocytes would be useful as a source of material for cell transplantation, bioartificial liver (BAL) support devices, studies on hepatocyte biology and hepatitis viruses and for drug testing.…”

“…Spontaneous differentiation of ES cells can be accomplished in serum-containing medium through the formation of embryoid bodies (EB)-containing cells representing ectoderm, mesoderm and endoderm lineages. Subsequent inoculation of the EB-derived cells on adherent matrix substrates allows spontaneous differentiation, producing occasional hepatocyte-like cells 3,6 . In vivo differentiation of mouse ES cells has also been accomplished by transplantation into the livers of carbon tetrachloride-treated mice.…”

In vitro differentiation of hepatic progenitor cells from mouse embryonic stem cells induced by sodium butyrate

“…Presently remarkable progress has been made in understanding the differentiation of ES cells into neural [Hancock et al, 2000], hematopoietic [Baron, 2001;Perlingeiro et al, 2001;Feng et al, 2005], endothelial , vascular [Sone et al, 2003;Yurugi-Kobayashi et al, 2003;Suzuki et al, 2005], and pancreatic stem/progenitor cells and then various mature cells. However, limited knowledge has been acquired for the differentiation of hepatic progenitor/stem cells, although hepatic differentiation from murine and human ES cells both in vitro and in vivo has been reported in recent studies [Hamazaki et al, 2001;Chinzei et al, 2002;Choi et al, 2002;Ishizaka et al, 2002;Jones et al, 2002;Miyashita et al, 2002;Yamada et al, 2002;Yin et al, 2002;Yamamoto et al, 2003;Kania et al, 2004;Ogawa et al, 2005;Teramoto et al, 2005;Teratani et al, 2005]. These hepatic cells from ES cells expressed some typical markers of mature hepatocytes and showed sufficient functions to rescue experimental liver injury when transplanted in vivo, which raised the hope to generate a transplantable cell source for the treatment of end-stage liver diseases.…”

Study of hepatocyte differentiation using embryonic stem cells

“…Spontaneous differentiation of mouse ES cells occurs when the cells are grown in suspension and form EBs [Abe et al, 1996;Chinzei et al, 2002;Jones et al, 2002;Miyashita et al, 2002;Yamada et al, 2002;Asahina et al, 2004;Kumashiro et al, 2005]. In order to direct the differentiation towards the endoderm lineage, various growth factors were used.…”