Differentiation of mouse embryonic stem cells into hepatocytes induced by a combination of cytokines and sodium butyrate

Zhou, Ming‐Ming; Li, Ping; Tan, Li; Qu, Su; Ying, Qi‐Long; Song, Houyan

doi:10.1002/jcb.22442

Cited by 26 publications

(20 citation statements)

References 48 publications

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“…Gene silencing associated with the aberrant methylation of promoter region of CpG islands is an acquired epigenetic alteration that serves as an alternative to genetic events in the inactivation of tumor suppressor and other genes in human cancers. NaBu has been applied in cellular and molecular research (23,24), particularly in antitumor research (25)(26)(27)(28). It plays a vital role in vitamin D-induced apoptosis through PTEN upregulation, which has potential benefit in gastric cancer therapy (29).…”

Section: Discussionmentioning

confidence: 99%

Sodium butyrate restores ASC expression and induces apoptosis in LS174T cells

Zhang

Bai

Ren

et al. 2012

International Journal of Molecular Medicine

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Abstract. Sodium butyrate (NaBu) is a short-chain fatty acid (SCFA), which has been proposed as a potential anticancer agent. Apoptosis-associated speck-like protein (ASC) is a pro-apoptotic signaling factor that is subjected to epigenetic silencing in human cancers. Modulation by the aberrant methylation of CpG islands of ASC is a well-characterized epigenetic mechanism, and the methylation-induced silencing of ASC has been observed in several types of tumors. NaBu induces cell cycle arrest, markers of cell differentiation and apoptosis in colon cancer. NaBu promotes transcriptional activation by relaxing the DNA conformation and displays anti-proliferative and differentiating activity in a wide variety of cancers. Thus, we used NaBu to investigate the relationship between the status of cell proliferation and the re-expression of ASC in colon carcinoma LS174T cells. Our experiments determined ASC re-expression at the protein level using western blotting. In addition, we used reverse transcriptionpolymerase chain reaction to detect the expression levels of ASC mRNA and an MTT assay to detect the inhibitory rate of cell growth. The apoptosis rate was also detected for further validation of the re-expression of ASC. The results showed that ASC re-expression was significantly increased in the LS174 cells following NaBu treatment in a time-and dose-dependent manner. The expression of ASC also induced the apoptosis of LS174T cells. These results suggest that NaBu plays a role in the reactivation of ASC expression and that the latter promotes the apoptosis of LS174T cells.

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Section: Discussionmentioning

confidence: 99%

Sodium butyrate restores ASC expression and induces apoptosis in LS174T cells

Zhang

Bai

Ren

et al. 2012

International Journal of Molecular Medicine

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“…Among these methods, the stepwise addition of two or more factors is most common. [19][20][21][22][23] However, if these complicated operations are not performed adequately, the death of ESCs can easily occur. In previous culture systems, remarkable reduction of the number of live cells was a technical obstacle to the induction of differentiation.…”

Section: Discussionmentioning

confidence: 99%

Differentiation of Monkey Embryonic Stem Cells to Hepatocytes by Feeder-Free Dispersion Culture and Expression Analyses of Cytochrome P450 Enzymes Responsible for Drug Metabolism

Maruyama

Matsunaga

Yamaori

et al. 2013

Biological & Pharmaceutical Bulletin

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We reported previously that monkey embryonic stem cells (ESCs) were differentiated into hepatocytes by formation of embryoid bodies (EBs). However, this EB formation method is not always efficient for assays using a large number of samples simultaneously. A dispersion culture system, one of the differentiation methods without EB formation, is able to more efficiently provide a large number of feeder-free undifferentiated cells. A previous study demonstrated the effectiveness of the Rho-associated kinase inhibitor Y-27632 for feeder-free dispersion culture and induction of differentiation of monkey ESCs into neural cells. In the present study, the induction of differentiation of cynomolgus monkey ESCs (cmESCs) into hepatocytes was performed by the dispersion culture method, and the expression and drug inducibility of cytochrome P450 (CYP) enzymes in these hepatocytes were examined. The cmESCs were successfully differentiated into hepatocytes under feeder-free dispersion culture conditions supplemented with Y-27632. The hepatocytes differentiated from cmESCs expressed the mRNAs for three hepatocyte marker genes (α-fetoprotein, albumin, CYP7A1) and several CYP enzymes, as measured by real-time polymerase chain reaction. In particular, the basal expression of cmCYP3A4 (3A8) in these hepatocytes was detected at mRNA and enzyme activity (testosterone 6β-hydroxylation) levels. Furthermore, the expression and activity of cmCYP3A4 (3A8) were significantly upregulated by rifampicin. These results indicated the effectiveness of Y-27632 supplementation for feeder-free dispersed culture and induction of differentiation into hepatocytes, and the expression of functional CYP enzyme(s) in cmESC-derived hepatic cells.Key words embryonic stem cell; differentiation; hepatocyte; monkey; cytochrome P450; feeder-free dispersed culture Investigation of drug metabolism with human hepatocytes is important in the early stages of drug development. However, primary human hepatocytes are short-lived and cannot be maintained in culture over the long term. In addition, there are large donor-dependent variations in drug metabolism. On the other hand, human embryonic stem cells (ESCs) are able to replicate infinitely and differentiate into various types of somatic cells including germ cells.1) Thus, they represent an attractive source to provide large numbers of cells that can be utilized for the development of candidate drug-screening strategies in place of primary cells.2) However, ethical and legal restrictions have limited the availability of human ESCs. The phenotype of human ESCs is known to closely resemble that of monkey ESCs but not mouse ESCs with regard to morphology, leukemia inhibitory factor responsiveness, gene expression profiles, and some disease models.1,3-6) Thus, monkey ESCs are a more suitable model for preclinical research of drug development. In particular, hepatocytes derived from monkey ESCs may be useful for pharmacokinetic studies, such as investigation of drug-drug interactions and the inducibility of drug-metabolizi...

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“…Cui combined epigenetic modification with cytokines gradually added to the cells in order to induce embryonic stem cells into hepatocytes using sodium butyrate and activin A treatment, achieving a production rate of functional liver cells of 10%-70% (Hay et al, 2008). The induction rate was improved through a novel three-step process with activin A, aFGF and sodium butyrate in recently reported work (Zhou et al, 2010). Sodium butyrate is a histone deacetylase inhibitor which promotes histone acetylation and opening of the chromatin structure; therefore a certain number of vital genes regulating cellular growth and terminal differentiation are activated for transcription (Zhou et al, 2010).…”

Section: Induction By Chromatin Modificationmentioning

confidence: 99%

“…The induction rate was improved through a novel three-step process with activin A, aFGF and sodium butyrate in recently reported work (Zhou et al, 2010). Sodium butyrate is a histone deacetylase inhibitor which promotes histone acetylation and opening of the chromatin structure; therefore a certain number of vital genes regulating cellular growth and terminal differentiation are activated for transcription (Zhou et al, 2010). Thus chromatin modulation is an important aspect of initiating gene transcription that impacts hepatocyte differentiation.…”

Section: Induction By Chromatin Modificationmentioning

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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Differentiation of mouse embryonic stem cells into hepatocytes induced by a combination of cytokines and sodium butyrate

Cited by 26 publications

References 48 publications

Sodium butyrate restores ASC expression and induces apoptosis in LS174T cells

Sodium butyrate restores ASC expression and induces apoptosis in LS174T cells

Differentiation of Monkey Embryonic Stem Cells to Hepatocytes by Feeder-Free Dispersion Culture and Expression Analyses of Cytochrome P450 Enzymes Responsible for Drug Metabolism

Directed hepatic differentiation from embryonic stem cells

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