Quantitative gene expression profiling reveals a fetal hepatic phenotype of murine ES-derived hepatocytes.

Jochheim, Andrea; Hillemann, Tina; Kania, Gabriela; Scharf, Jennifer; Attaran, Masoumeh; Manns, Michael P.; Wobus, Anna M.; Ott, Michael

doi:10.1387/ijdb.15005571

Cited by 57 publications

(46 citation statements)

References 28 publications

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“…However, although these markers are widely used to assess function of mature hepatocytes isolated from adult livers, they may also be expressed in nonhepatic lineage embryonic cells. [14][15][16][17][18][19] In addition, other markers often used in evaluating hepatocyte differentiation, such as glucose-6-phosphatase, have also been found, using reverse transcription-polymerase chain reaction (RT-PCR), in cells of a variety of other tissues including pancreatic islets. 28 In general, although RT-PCR is a powerful tool, only a limited number of genes are generally evaluated, and many of these genes are expressed in a variety of embryonic and adult tissues.…”

Section: Discussionmentioning

confidence: 99%

“…Two micrograms of total cellular RNA was reverse-transcribed from a "capture-sequence"-containing oligod(T) 18 primer using SuperScript II (Invitrogen). After al kaline hydrolysis and neutralization, the cDNA target was hybridized with probes on the array at 58°C for 12 h using a Ventana Discovery workstation (Ventana Medical Systems, Tuscon, AZ) and washed to high stringency.…”

Section: Hybridizationmentioning

confidence: 99%

“…10 or cells that express genes for albumin (ALB), alpha-feto protein (AFP), and transthyretin protein (TRT). [14][15][16][17][18] Spontaneous differentiation of human EBs into hepatocyte lineage cells, in the absence of both ECM and exogenous growth factor supplementation, has also been identified using gene microarray analysis of several known mature liver specific proteins. 20,21 Despite the growing number of studies reporting ES/hepatocyte differentiation, isolation of pure populations of neither committed hepatocyte precursors nor fully differentiated hepatocytes have been reported.…”

Section: Introductionmentioning

confidence: 99%

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Embryoid Body–Mediated Differentiation of Mouse Embryonic Stem Cells Along a Hepatocyte Lineage: Insights from Gene Expression Profiles

et al. 2006

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Pluripotent embryonic stem (ES) cells represent a promising renewable cell source for the generation of functional differentiated cells. Previous studies incorporating embryoid body (EB)-mediated stem cell differentiation have, either spontaneously or after growth factor and extracellular matrix protein supplementation, yielded populations of hepatocyte lineage cells expressing mature hepatocyte markers such as albumin (ALB). In an effort to promote ES cell commitment to the hepatocyte lineage, we have evaluated the effects of four culture conditions on albumin and gene expression in differentiating ES cells. Quantitative in situ immunofluorescence and cDNA microarray analyses were used to describe not only lineage specificity but also to provide insights into the effects of disparate culture environments on the mechanisms of differentiation. The results of these studies suggest that spontaneous and collagen-mediated differentiation induce cells with the highest levels of ALB expression but mature liver specific genes were only expressed in the spontaneous condition. Further analysis of gene expression profiles indicated that two distinct mechanisms may govern spontaneous and collagen-mediated differentiation.

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Section: Discussionmentioning

confidence: 99%

Section: Hybridizationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Embryoid Body–Mediated Differentiation of Mouse Embryonic Stem Cells Along a Hepatocyte Lineage: Insights from Gene Expression Profiles

et al. 2006

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“…Basma et al were able to obtain functional hepatocytes from human ESCs using a similar protocol (Basma et al, 2009 this method with additional factors such as BMP4, FGF10 and retinoic acid, and they produced human hepatocytes exhibiting hepatic functions including glycogen storage, cytochrome activity and low-density lipoprotein uptake (Touboul et al, 2010). There are numerous factors in serum such as hormones and growth factors, which regulate the process of directed differentiation into hepatocytes and yet increase uncertainty of differentiation process at the same time (Jochheim et al, 2004;Saito et al, 2006). Thus many studies focused on serum-free conditions of induction (Agarwal et al, 2008;Hay et al, 2008;Shiraki et al, 2008;Zamule et al, 2011).…”

Section: Soluble Factor-induced Methodsmentioning

confidence: 99%

Directed hepatic differentiation from embryonic stem cells

Chen

Zeng

2011

Protein Cell

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The liver is the largest internal organ in mammals, and is important for the maintenance of normal physiological functions of other tissues and organs. Hepatitis, cirrhosis, liver cancer and other chronic liver diseases are serious threats to human health, and these problems are compounded by a scarcity of liver donors for transplantation therapies. Directed differentiation of embryonic stem cells to liver cells is a promising strategy for obtaining hepatocytes that can be used for cell transplantation. In vitro hepatocyte differentiation of embryonic stem cells requires a profound understanding of normal development during embryonic hepatogenesis. Here we provide a simple description of hepatogenesis in vivo and discuss directed differentiation of embryonic stem cells into hepatocytes in vitro.

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“…In order to direct the differentiation towards the endoderm lineage, various growth factors were used. In most of the protocols the cells were grown as EBs for several days and then plated on adherent matrix and treated with several growth factors [Hamazaki et al, 2001;Kuai et al, 2003;Hu et al, 2004;Imamura et al, 2004;Jochheim et al, 2004;Kania et al, 2004;Shirahashi et al, 2004;Teratani et al, 2005]. Usually, collagen was chosen as the matrix for growing the cells since the liver bud proliferates and migrate into the septum transversum mesenchyme that is composed of loose connective tissue containing collagen.…”

Section: Embryonic Stem Cells and Hepatocytesmentioning

confidence: 99%

Study of hepatocyte differentiation using embryonic stem cells

Lavon

Benvenisty

2005

J of Cellular Biochemistry

114

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The liver has many crucial functions including metabolizing dietary molecules, detoxifying compounds, and storing glycogen. The hepatocytes, comprising most of the liver organ, progressively modify their gene expression profile during the fetal development according to their roles in the different phases of development. Embryonic stem (ES) cells serve as a major tool in understanding liver development. These cells may also serve as a source of hepatic cells for cellular therapy. In this review, we aim to summarize the research that has been performed in the field of hepatocyte differentiation from mouse and human ES cells. We discuss the various methodologies for the differentiation of ES cells towards hepatic cells using either spontaneous or directed differentiation protocols. Although many protocols for differentiating ES cells to hepatic cells have been developed, the analysis of their status is not trivial and can lead to various conclusions. Hence, we discuss the issues of analyzing hepatocytes by means of the specificity of the markers for hepatocytes and the status of the cells as fetal or adult hepatocytes.

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Quantitative gene expression profiling reveals a fetal hepatic phenotype of murine ES-derived hepatocytes.

Cited by 57 publications

References 28 publications

Embryoid Body–Mediated Differentiation of Mouse Embryonic Stem Cells Along a Hepatocyte Lineage: Insights from Gene Expression Profiles

Embryoid Body–Mediated Differentiation of Mouse Embryonic Stem Cells Along a Hepatocyte Lineage: Insights from Gene Expression Profiles

Directed hepatic differentiation from embryonic stem cells

Study of hepatocyte differentiation using embryonic stem cells

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