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

Soto-Gutiérrez, Alejandro; Navarro-Alvarez, Nalú; Zhao, Debiao; Rivas‐Carrillo, Jorge David; Lebkowski, Jane; Tanaka, Noriaki; Fox, Ira J.; Kobayashi, Naoya

doi:10.1038/nprot.2007.18

Cited by 120 publications

(94 citation statements)

References 17 publications

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“…31) EBs formed by culture for 2 d can be successfully induced to differentiate into definitive endoderm by culture in activin A and FGF-2. 32) We do not know the reason why the expression levels of hepatocyte marker genes, especially ALB, in the cells cultured for 2 d for EB formation from cmES cells was higher than those in cells cultured for 5 d (Fig. 3).…”

Section: Discussionmentioning

confidence: 97%

Differentiation of Monkey Embryonic Stem Cells into Hepatocytes and mRNA Expression of Cytochrome P450 Enzymes Responsible for Drug Metabolism: Comparison of Embryoid Body Formation Conditions and Matrices

Momose

Matsunaga

Murai

et al. 2009

Biological & Pharmaceutical Bulletin

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Examination of drug metabolism using 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. Considerable donor-dependent variations are also problematic. Human embryonic stem (ES) cells are pluripotent cells derived from the inner cell mass of blastocysts and are capable of differentiating into three embryonic germ layers and germ cells.1) The cells apparently differentiate into various types of mature cells, and are thereby an attractive source for routine access to large numbers of cells that can be used for the development of candidate drug-screening strategies replacing primary cells.2) However, ethical considerations have limited the availability of human ES cells. The phenotype of human ES cells is known to be similar to that of monkey ES cells but differs from that of mouse ES cells with regard to morphology, response to leukemia inhibitory factor, and gene expression patterns. 1,[3][4][5] Research using monkey ES cells is considered to be useful for investigation of differentiation mechanisms in primate ES cells and models of human ES cells. Recently, it has been shown that monkey ES cells can be differentiated into various cell types-including neurons, hematopoietic cells, and pancreatic cells-using growth factors. [6][7][8] Hepatocytes derived from monkey ES cells may be useful for pharmacokinetic examinations such as induction of drug metabolism enzymes and interactions of candidate drugs. To date however, there have been few reported studies describing the differentiation of monkey ES cells into hepatocytes. 9,10) After the emergence of the liver bud from the developing gut tube, the level of hepatic maturation is characterized by the expression of liver-and stage-specific genes. For example, alfa-fetoprotein (AFP) is an early hepatic marker, expressed by hepatoblasts in the liver bud until birth.11,12) The synthesis of AFP decreases dramatically after birth and only trace amounts are expressed in the adult liver. In contrast, albumin (ALB), the most abundant protein synthesized by hepatocytes, is initially expressed at lower levels in early fetal hepatocytes but this increases as the hepatocytes mature, reaching a maximum in adult hepatocytes. 13,14) The mRNA expression of cytochrome P450 7A1 (CYP7A1) which is a rate-limiting enzyme in the conversion of cholesterol to bile acids in liver 15) is detected in fetal liver of third trimester of pregnancy.16) After birth, CYP7A1 increases several-fold with age both at the enzyme activity level and the mRNA level. 17,18) The in vitro approaches involve the formation of embryoid bodies (EBs) to mimic the inductive microenvironment required for liver organogenesis [19][20][21] and treatment with specific growth factors and cytokines critical for hepatic differentiation.22) At present, culture systems for ES cells have mainly used gelatin-or collagen-coated plates as the matrix for the maintenance of cells in an undifferentiated state an...

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Section: Discussionmentioning

confidence: 97%

Differentiation of Monkey Embryonic Stem Cells into Hepatocytes and mRNA Expression of Cytochrome P450 Enzymes Responsible for Drug Metabolism: Comparison of Embryoid Body Formation Conditions and Matrices

Momose

Matsunaga

Murai

et al. 2009

Biological & Pharmaceutical Bulletin

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“…4K). Third, using established methods for directed differentiation of mouse ES cells, we examined whether or not rat limbal iPSc are amenable to differentiation into functional neurons [44], cardiomyocytes [19], and hepatocytes [21] as described below (Fig. 5).…”

Section: Resultsmentioning

confidence: 99%

“…Rat limbal iPSCs were differentiated into hepatocytes by a previously described method [21]. Briefly, EBs generated by rat limbal iPSCs were cultured in matrigel (BD Bioscience)-coated dish in differentiation medium I [DMEM/F12, 1% FBS, 1% nonessential amino acids, 1% nucleosides, 1% penicillin þ streptomycin, 1% glutamic acid, 3% BSA, 100 ng/ml of FGF2, and 100 ng/ml of Activin A (R&D Systems)] for 3 days at 37 C (induction phase).…”

Section: Methodsmentioning

confidence: 99%

Non Cell-Autonomous Reprogramming of Adult Ocular Progenitors: Generation of Pluripotent Stem Cells without Exogenous Transcription Factors

et al. 2009

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Direct reprogramming of differentiated cells to induced pluripotent stem (iPS) cells by ectopic expression of defined transcription factors (TFs) represents a significant breakthrough towards the use of stem cells in regenerative medicine (Takahashi and Yamanaka Cell 2006;126:663–676). However, the virus‐mediated expression of exogenous transcription factors could be potentially harmful and, therefore, represents a barrier to the clinical use of iPS cells. Several approaches, ranging from plasmid‐mediated TF expression to introduction of recombinant TFs (Yamanaka Cell 2009;137:13–17; Zhou, Wu, Joo et al. Cell Stem Cell 2009;4:381–384), have been reported to address the risk associated with viral integration. We describe an alternative strategy of reprogramming somatic progenitors entirely through the recruitment of endogenous genes without the introduction of genetic materials or exogenous factors. To this end, we reprogrammed accessible and renewable progenitors from the limbal epithelium of adult rat eye by microenvironment‐based induction of endogenous iPS cell genes. Non cell‐autonomous reprogramming generates cells that are pluripotent and capable of differentiating into functional neurons, cardiomyocytes, and hepatocytes, which may facilitate autologous cell therapy to treat degenerative diseases. STEM CELLS 2009;27:3053–3060

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“…Impressive studies have been published by SatoGutierrez et al (2006). Their differentiation protocol (specifically described by Sato-Gutierrez et al, 2007) utilized EB formation and then treatment with Activin A and FGF2. After that, it required indirect co-culturing with human liver non-parenchymal cell lines (cholangiocytes, stellate cells, and liver endothelial cells) and treatment with HGF, DMSO, and DEX (Sato-Gutierrez et al, 2006).…”

Section: Selection Of Es Cell-derived Hepatic Progenitors Bmp-4 Has mentioning

confidence: 99%

Stem cell plasticity: Learning from hepatogenic differentiation strategies

Banas¹,

Yamamoto

Teratani³

et al. 2007

Developmental Dynamics

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Many studies on stem cell plasticity are challenging the concept that stem cells contain an intrinsically predefined, unidirectional differentiation program. This means that the developmental fate of a stem cell is dependent on the general potential of the cell (pre-determined stem cell fate) as well as on microenvironmental cues, such as stimuli from growth factors (stem cell niche). Here, we reviewed reports that examined the hepatocyte differentiation ability of stem cells from two different sources: embryonic stem cells and adult stem cells. All of those stem cells revealed the ability to give rise to hepatocyte-like cells using different induction strategies. However, it is still not clear which of those stem cells would be the best source for hepatocyte replacement or which would be the best protocol. We herein present the current knowledge regarding available protocols and factors used in order to obtain functional hepatocytes from stem cells. Developmental Dynamics 236:3228 -3241, 2007.

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

Cited by 120 publications

References 17 publications

Differentiation of Monkey Embryonic Stem Cells into Hepatocytes and mRNA Expression of Cytochrome P450 Enzymes Responsible for Drug Metabolism: Comparison of Embryoid Body Formation Conditions and Matrices

Differentiation of Monkey Embryonic Stem Cells into Hepatocytes and mRNA Expression of Cytochrome P450 Enzymes Responsible for Drug Metabolism: Comparison of Embryoid Body Formation Conditions and Matrices

Non Cell-Autonomous Reprogramming of Adult Ocular Progenitors: Generation of Pluripotent Stem Cells without Exogenous Transcription Factors

Stem cell plasticity: Learning from hepatogenic differentiation strategies

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