growth and differentiation of human hepatocytes

use in a bioartificial liver system by Christine Vynetta Burt

Publisher: University of Birmingham in Birmingham

Written in English
Published: Pages: 176 Downloads: 781
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Edition Notes

Thesis (Ph.D) - University of Birmingham, School of Biosciences, Faculty of Science.

Statementby Christine Vynetta Burt.
The Physical Object
Pagination176 p., 12 leaves of coloured plates :
Number of Pages176
ID Numbers
Open LibraryOL18805224M

hepatocytes derived from human embryonic stem cells (hESCs) are anticipated to become a potentially useful cell source for various cell therapies, bioartificial livers, and drug discovery support systems. Although several articles have reported the differentiation of hESCs into hepatocyte-like cells in vitro by various methods (2, 5, 6, 16), it remains unknown which extracellular matrixes. Hepatocyte growth factor was found to play a positive role in promoting hepatocyte differentiation. Our differentiation system has shown that hESCs can be differentiated to hepatocyte-like cells capable of executing a range of hepatocyte functions. Therefore, it presents a proof-of-principle of potential applications of using the hESC-derived. CONCLUSION We have developed an efficient way to direct the differentiation of human embryonic stem cells into cells that exhibit characteristics of mature hepatocytes. Our studies should facilitate searching the molecular mechanisms underlying human liver development, and form the basis for hepatocyte transplantation and drug tests.   New Approaches in the Differentiation of Human Embryonic Stem Cells and Induced Pluripotent Stem Cells toward Hepatocytes 19 February | Stem Cell Reviews and Reports, Vol. 7, No. 3 Hepatic Differentiation of Human Embryonic Stem Cells Is Promoted by Three-Dimensional Dynamic Perfusion Culture Conditions.

  Maintenance of liver-specific function of hepatocytes in culture is still difficult. Improved culture conditions may enhance the cell growth and function of cultured cells. We investigated the effect of three-dimensional culture under flow conditions, and the influence of surface modifications in hepatocyte cultures.   Introduction. L iver cell therapies, including hepatocyte transplantation and the use of bioartificial liver systems, have been used to reestablish liver function and to act as a bridge to whole-liver transplantation for patients with hereditary liver disease or fulminant liver failure. 1,2 The major limitation of these approaches is the availability of human livers as a source of whole-liver. Abstract. Hepatocytes derived from human embryonic stem cells (ESCs) are a potential cell source for regenerative medicine. However, the successful differentiation of human ESCs into mature hepatocytes has been difficult to achieve because the definitive mechanisms governing hepatocyte differentiation have not yet been well defined. Growth Factors, Chemicals and Antibodies. Ac-tivin A, acidic FGF (aFGF), basic FGF (bFGF), FGF4, hepatocyte growth factor (HGF), and bone morphoge-netic protein-2 (BMP2) were purchased from Peprotech (Rocky Hill, NJ), Oncostatin M (OSM), human BMP4 and recombinant mouse Noggin were purchased from R&D System (Minneapolis, MN). Dexamethasone.

The in vitro toxicology field seeks for reliable human relevant hepatic models for predicting xenobiotics metabolism and for the safety assessment of chemicals and developing drugs. The low availability and rapid loss of the phenotype or low biotransformation activity of primary hepatocytes urged the stem cell differentiation into hepatocyte-like cells (HLCs). For further differentiation, the specified hepatic progenitor cells on the R-Fc-coated surface were treated with hepatocyte growth factor for an additional 5 days, and expression of AFP and HNF4A was detected by immunocytochemistry. Fig 6A shows that more than 90% of cells expressed HNF4A, and that HNF4A-positive cells co-expressed AFP at day The generated liver organoids exhibited favorable growth and differentiation of hepatocytes and cholangiocytes, recapitulating the key features of human liver formation with cellular heterogeneity. Impact Statement. Although oxygen is a vital nutrient for the hepatocytes in vitro, few reports have focused on its effect during hepatic differentiation of induced pluripotent stem cells (iPSCs).In this report, we performed the hepatic differentiation of human iPSCs (hiPSCs) under different atmospheric oxygen concentrations and oxygen supply fluxes to investigate the effects of oxygen in.

growth and differentiation of human hepatocytes by Christine Vynetta Burt Download PDF EPUB FB2

Growth and differentiation of colony-forming human hepatocytes in vitro Author links open overlay panel Chihiro Yamasaki 1 2 Chise Tateno 1 2 3 Akio Aratani 2 Chimoto Ohnishi 1 2 Shigeru Katayama 2 4 Toshihiko Kohashi 4 Hiroshi Hino 2 4 Hiroyuki Marusawa 5 Toshimasa Asahara 3 4 Katsutoshi Yoshizato 1 2 3 6Cited by: With the development of regeneration medicine, many researchers have attempted hepatic differentiation from nonhepatic‐origin cell sources.

The differentiation of embryonic stem (ES) cells into Cited by:   1. Introduction. Generally, hepatocytes in vitro can be maintained as replicating differentiated cells for a limited period. We devised a culture medium, hepatocyte clonal growth medium (HCGM), that supports the growth of rat hepatocytes for a longer period.Using HCGM we were able to demonstrate the presence of the colony-forming parenchymal hepatocytes (CF-PHs) as small hepatocytes Cited by: The availability of the isolated hepatocyte preparation as cells in suspension or culture has undoubtedly facilitated research on the liver.

This was emphasised in our book, published (with Dr. Greg Barritt) inwhich described in detail methods of preparation and the properties of the isolated hepatocytes.

This was emphasised in our book, published (with Dr. Greg Barritt) inwhich described in detail methods of preparation and the properties of the isolated hepatocytes. It also discussed the usefulness of the preparation for the study of intermediary and xenobiotic metabolism, calcium ion transport, and the growth and differentiation of hepatocytes in culture.

The first part of the book focuses on the use of these particular liver-based in vitro models to study the different aspects of the hepatocyte life cycle, including cell growth, differentiation and cell death.

Primary human hepatocytes are still the gold standard, but they have substantial disadvantages such as rapid dedifferentiation in vitro and lack of cell proliferation.

In addition to primary human hepatocytes, liver cancer-derived cell lines such as HepG2, cytochrome P (CYP) overexpressing HepG2 cell clones and HepaRG were studied.

Protocol for directed differentiation of human pluripotent stem cells toward a hepatocyte fate that facilitate the reproducible differentiation of hepatocytes from a wide-range of hESCs and hiPSCs (Delaforest et al., ; Si-Tayeb et al., b).

Although the basic protocol described here is efficient and has proven effective in. Abstract. AIM: Clinical application of human hepatocytes (HC) is hampered by the progressive loss of growth and differentiation in vitro. The object of the study was to evaluate the effect of a biphasic culture technique on expression and activation of growth factor receptors and differentiation of human adult HC.

Differentiation of mouse embryonic stem cells to hepatocyte-like cells by co-culture with human liver nonparenchymal cell lines. Nat Protoc 2, – Stafford, D. Hornbruch, A.

Mueller, P.R. Prince, V.E. ave been shown to be useful as scaffolds for seeding and culturing various types of cells. In this study, a porous sponge scaffold of modified PLGA polymer with collagen was investigated for its ability to improve the growth and metabolism of human hepatocytes.

We evaluated the biocompatibility of collagen-modified PLGA (C-PLGA) scaffolds with hepatocytes isolated from human liver.

Cell. However, all directed differentiation protocols for hepatocyte-like cells (HLCs) published to date have relied on the use of recombinant growth factors such as activin A, Wnt3a, hepatocyte growth factor (HGF), oncostatin M (OSM), fibroblast growth factor 4 (FGF4), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), and bone morphogenetic protein 4 (BMP4).

Differentiation of Human Parthenogenetic Embryonic Stem Cells into Functional Hepatocyte-like Cells. Organogenesis: Vol. 16, No. 4, pp.

Muscle progenitor cell isolation, growth, and seeding. Primary human skeletal muscle cells isolated by needle biopsy [] were expanded in myoblast growth medium [MGM; SkGM (Cambrex Bio Science, Walkersville, MD) plus 15% (v/v) fetal bovine serum].Biopsies were performed on adult volunteers according to procedures approved by the Institutional Clinical Review Board of the Miriam Hospital.

Gerbal-Chaloin S. et al. () Isolation and Culture of Adult Human Liver Progenitor Cells: In Vitro Differentiation to Hepatocyte-Like Cells. In: Maurel P. (eds) Hepatocytes. Methods in Molecular Biology (Methods and Protocols), vol Currently, most of these strategies employ step-wise differentiation approaches with recombinant growth-factors or small-molecule analogs to recapitulate developmental signaling pathways.

Here, we tested the efficacy of a small-molecule based differentiation protocol for the generation of hepatocyte-like cells from human pluripotent stem cells.

Abstract. Human embryonic stem cells (hESCs) can be progressively differentiated into definitive endoderm (DE), hepatic progenitors, and hepatocytes, and thus provide an excellent model system for the mechanistic study of hepatocyte differentiation, which is currently poorly understood.

Here, we found that insulin enhanced hepatocyte differentiation from hESC-derived DE. Abstract. Hepatocyte growth factor (HGF) is a paracrine factor involved in organogenesis, tissue repair, and wound healing.

We report here that HGF promotes osteogenic differentiation through the transcription of key osteogenic markers, including osteocalcin, osterix, and osteoprotegerin in human mesenchymal stem cells and is a necessary component for the.

Katsura et al. reported that primary human hepatocytes were cultured in keratinocyte-stimulating factor medium supplemented with 10% human serum, 10 mM nicotinamide, 10 ng/ml epidermal growth. Background: The advent of human-induced pluripotent stem cells holds great promise for producing ample individualized hepatocytes.

Although previous efforts have succeeded in generating hepatocytes from human pluripotent stem cells in vitro by viral-based expression of transcription factors and/or addition of growth factors during the differentiation process, the safety issue of viral transduction.

Methods: To generate human hepatocytes, human embryonic stem cells were differentiated by sequential culture in fibroblast growth factor 2 and human activin-A, hepatocyte growth factor, and dexamethasone. Functional hepatocytes were isolated by sorting for. Several reports have been published on the differentiation of human iPS cells into hepatocyte-like cells; however, the cells were insufficient for application in drug metabolism studies.

In this study, we aimed to establish effective methods for differentiation of human iPS cells into hepatocytes. Two human iPS cell lines were differentiated by addition of activin A, dimethyl sulfoxide, hepatocyte growth factor.

The directed differentiation of human embryonic stem cells (hESCs) or human induced pluripotent stem cells (hiPSCs) into hepatocytes could facilitate a rational study of the molecular mechanisms underlying human liver development as well as provide a renewable source of exogenous hepatocytes for drug toxicity testing and cell-based therapeutics.

Moreover, if hepatocytes were produced from. Abstract. Directing embryonic stem cell (ESC)-derived hepatocytes is critical in understanding hepatic differentiation and applying cell-based treatment to severe liver diseases. While growth factor-based strategies are widely used, using chemical cues could present an alternative to optimize the strategies for stem cell differentiation.

A three‐step protocol to differentiate human embryonic stem cells (hESCs) to hepatocytes. (A) Schematic representation illustrating the three‐stage procedure in differentiation of hESCs to hepatocytes.(B) Images showing sequential morphological changes from hESCs (H7) to hepatocytes through the definitive endoderm and hepatoblast during differentiation.

Hepatocyte growth factor (HGF) can influence epithelial cell growth and differentiation. We examined the actions of recombinant human HGF (rhHGF) on the differentiation of human primary tracheal epithelial (HTE) cells cultured in vitro for up to 96 h. Basolateral, but not apical, treatment of confluent HTE cell sheets for 48 h with rhHGF led to.

Kazuo Takayama, Establishment of a Method of Hepatocyte Differentiation from Human Pluripotent Stem Cells for Innovative Drug Developmentヒト多能性幹細胞由来肝細胞を用いた次世代型創薬の実現を目指した基盤技術創成, YAKUGAKU ZASSHI, /yakushi,10, (), ().

Some examples of these are; Liebovitz L, DMEM/F, RPMIWaymouth's MB /1 and Williams Medium E. Growth of hepatocytes in these media requires the use of of serum. Our most advanced formulation for culturing primary, secondary, and cloned immortalized epithelial hepatocytes is Hwhich is an enriched variation of Leibovitz L.

Mature adult parenchymal hepatocytes, typically of restricted capacity to proliferate in culture, can now enter into clonal growth under the influence of hepatocyte growth factor (scatter factor) (HGF/SF), epidermal growth factor (EGF), and transforming growth factor alpha (TGFalpha) in the presence of a new chemically defined medium (HGM).

Methods. To generate human hepatocytes, human embryonic stem cells were differentiated by sequential culture in fibroblast growth factor 2 and human activin-A, hepatocyte growth factor, and dexamethasone. Functional hepatocytes were isolated by sorting for.

Human hepatocyte-specific markers were detected in the liver 3 h after injection, but the cells did not resemble hepatocyte-like cells. However, by 2 and 7 days after injection, hepatocyte-like cells expressing CK18, AFP, ALB, and TPH2 were found.

The levels of hepatocyte-specific markers increased in a time-dependent manner.HepaRG cells are liver bipotent progenitors acquiring hepatocytes features when differentiated in the presence of dimethylsulfoxide (DMSO). Differentiated HepaRG (dHepaRG) are considered the best surrogate model to primary human hepatocytes (PHH) and are susceptible to several hepatotropic viruses, including Hepatitis B Virus (HBV) and Hepatitis Delta Virus (HDV) infection.Other articles where Hepatocyte is discussed: digestive system disease: Liver: the three functional components: the hepatocyte (liver cell), the bile secretory (cholangiolar) apparatus, or the blood vascular system.

Although an agent tends to cause initial damage in only one of these areas, the resulting disease may in time also involve other components.

Thus, although viral hepatitis.