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            , ESC‐Derived Cells Engraft and Differentiate into Hepatocytes in Vivo

Yin, Yijun; Lim, Y.C.; Salto–Tellez, Manuel; Ng, Soon-Chye; Lin, Chyuan‐Sheng; Lim, Sai‐Kiang

doi:10.1634/stemcells.20-4-338

Cited by 80 publications

(66 citation statements)

References 16 publications

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“…In the present study, ES cell-derived GFP ϩ cells isolated with high purity using FACS were subjected to normal growth regulation and did not produce teratomas within 104 days after transplantation into the diseased liver of MUP-uPA/SCID mice, whereas undifferentiated ES cells gave rise to numerous tumors only 10 days after transplantation. Similar results were observed after transplantation of GFP-positive 12,26 or PECAM1-negative cells isolated from EB-derived cells. 7 Thus, differentiation of ES cells in culture and purification using either positive or negative selection may significantly decrease the tumorigenic potential of ES cells.…”

Section: Discussionsupporting

confidence: 82%

“…Although several studies have shown that ES cell-derived hepatocytes incorporate into recipient liver upon transplantation 7,10,12,26 the repopulation efficiency was very low, not exceeding 0.1%. 12,26 In our study, ES cell-derived hepatocytes replaced from 1% to 20% of the parenchyma in randomly selected areas of the liver in MUPuPA/SCID mice. The repopulating ES cell-derived cells displayed characteristics of normal differentiated hepatocytes and established structural connections with each other and with neighboring host hepatocytes, indicating their functional integration.…”

Section: Discussionmentioning

confidence: 98%

“…ALB promoter has an obvious advantage over the alpha-fetoprotein promoter used in previous studies 16,26 because it can trace mature hepatocytes as well as hepatocyte precursors. 27 In our study, the expression of the GFP transgene was stable and did not decrease during ES cell growth, as reported previously.…”

Section: Discussionmentioning

confidence: 99%

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Hepatic precursors derived from murine embryonic stem cells contribute to regeneration of injured liver

et al. 2006

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We established an efficient system for differentiation, expansion and isolation of hepatic progenitor cells from mouse embryonic stem (ES) cells and evaluated their capacity to repopulate injured liver. Using mouse ES cells transfected with the green fluorescent protein (GFP) reporter gene regulated by albumin (ALB) enhancer/promoter, we found that a serum-free chemically defined medium supports formation of embryoid bodies (EBs) and differentiation of hepatic lineage cells in the absence of exogenous growth factors or feeder cell layers. The first GFP ؉ cells expressing ALB were detected in close proximity to "beating" myocytes after 7 days of EB cultures. GFP ؉ cells increased in number, acquired hepatocyte-like morphology and hepatocytespecific markers (i.e., ALB, AAT, TO, and G6P), and by 28 days represented more than 30% of cells isolated from EB outgrowths. The FACS-purified GFP ؉ cells developed into functional hepatocytes without evidence of cell fusion and participated in the repairing of diseased liver when transplanted into MUP-uPA/SCID mice. The ES cell-derived hepatocytes were responsive to normal growth regulation and proliferated at the same rate as the host hepatocytes after an additional growth stimulus from CCl 4 -induced liver injury. The transplanted GFP ؉ cells also differentiated into biliary epithelial cells. In conclusion, a highly enriched population of committed hepatocyte precursors can be generated from ES cells in vitro for effective cell replacement therapy. Supplementary material for this article can be found on the HEPATOLOGY website H epatocyte transplantation is an effective treatment for liver failure and/or end-stage liver disease. However, the shortage of donor organs and the difficulties of cyropreservation and long-term culturing of mature hepatocytes have limited the clinical application of cell-based therapy. Recently, the use of embryonic stem (ES) cells has attracted considerable interest for cell replacement therapy because of their capacity to proliferate indefinitely in vitro while retaining the potential to differentiate into all types of cells including hepatocytes. [1][2][3][4][5] Clinical application of ES cells requires a simple and efficient protocol for directing their differentiation into a specific cell type. Several studies have reported the induction of ES cell differentiation into hepatocytes both in vitro 6-9 and in vivo. 10 The in vitro approaches involve the formation of embryoid bodies (EBs) to re-create the inductive microenvironment required for liver organogenesis 11,12 or treatment with specific growth factors and cytokines critical for hepatocyte differentiation such as hepatocyte growth factor, fibroblast growth factor, and oncostatin. 1,13 Also, the introduction of genes promoting endodermal differentiation into ES cells, 14 modification of culture microenvironment by supplementation with extracellular matrix proteins or coculture with other cell types, 15,16 is used to direct ES cells toward a hepatocytic lineage. Nevertheless, these strategie...

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Section: Discussionsupporting

confidence: 82%

Section: Discussionmentioning

confidence: 98%

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Hepatic precursors derived from murine embryonic stem cells contribute to regeneration of injured liver

et al. 2006

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“…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.…”

mentioning

confidence: 99%

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

Zhou

Xiang

Shao

et al. 2006

J of Cellular Biochemistry

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Recently it was shown that embryonic stem (ES) cells could differentiate into hepatocytes both in vitro and in vivo, however, prospective hepatic progenitor cells have not yet been isolated and characterized from ES cells. Here we presented a novel 4-step procedure for the differentiation of mouse ES cells into hepatic progenitor cells and then hepatocytes. The differentiated hepatocytes were identified by morphological, biochemical, and functional analyses. The hepatic progenitor cells were isolated from the cultures after the withdrawal of sodium butyrate, which was characterized by scant cytoplasm, ovoid nuclei, the ability of rapid proliferation, expression of a series of hepatic progenitor cell markers, and the potential of differentiation into hepatocytes and bile duct-like cells under the proper conditions that favor hepatocyte and bile epithelial differentiation. The differentiation of hepatocytes from hepatic progenitor cells was characterized by a number of hepatic cell markers including albumin secretion, upregulated transcription of glucose-6-phophatase and tyrosine aminotransferase, and functional phenotypes such as glycogen storage. The results from our experiments demonstrated that ES cells could differentiate into a novel bipotential hepatic progenitor cell and mature into hepatocytes with typical morphological, phenotypic and functional characteristics, which provides an useful model for the studies of key events during early liver development and a potential source of transplantable cells for cell-replacement therapies.

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“…While promising, the embryonic stem cell derivates have shown a low engraftment efficiency, lack of function in the liver [16] [17], and even fusion of the injected cells to the surrounding hepatocytes [18]. Therefore, a more direct approach based on the developmental relationship between ES cells, definitive endoderm and hepatocellular function is required in order to increase efficiency, function and safety of ES cell derivate transplants into the liver.…”

Section: Introductionmentioning

confidence: 99%

Detection and Characterization of Hepatic Engraftment of Embryonic Stem Derived Cells by Fluorescent Stereomicroscopy

Caballero

Lightfoot

Lapaglia

et al. 2007

Journal of Surgical Research

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Background-Embryonic Stem (ES) cells have been investigated as a potential replacement therapy for failed organs, such as the liver. However, detection of hepatic engraftment from candidate stem cells has been difficult due to low engraftment efficiency. Previous detection methods required that the graft be processed by molecular and/or immunohistochemical techniques, limiting further functional studies. This study evaluated the use of tri-dimensional (3-D) fluorescent stereomicroscopy for gross detection of ES cell derived hepatic engraftment.

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AFP ⁺ , ESC‐Derived Cells Engraft and Differentiate into Hepatocytes in Vivo

Cited by 80 publications

References 16 publications

Hepatic precursors derived from murine embryonic stem cells contribute to regeneration of injured liver

Hepatic precursors derived from murine embryonic stem cells contribute to regeneration of injured liver

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

Detection and Characterization of Hepatic Engraftment of Embryonic Stem Derived Cells by Fluorescent Stereomicroscopy

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AFP + , ESC‐Derived Cells Engraft and Differentiate into Hepatocytes in Vivo

Cited by 80 publications

References 16 publications

Hepatic precursors derived from murine embryonic stem cells contribute to regeneration of injured liver

Hepatic precursors derived from murine embryonic stem cells contribute to regeneration of injured liver

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

Detection and Characterization of Hepatic Engraftment of Embryonic Stem Derived Cells by Fluorescent Stereomicroscopy

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AFP ⁺ , ESC‐Derived Cells Engraft and Differentiate into Hepatocytes in Vivo