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Hepatocytes obtained from mouse GPSCs are functional and show similar expression profiles to fetal primary hepatocytes

ABSTRACT: Germline cell-derived pluripotent stem cells (GPSCs) are similar to embryonic stem (ES) cells in that they can proliferate intensively and differentiate into a variety of cell types, including cardiomyocytes and neurons. In this report, mouse GPSCs were induced to differentiate into hepatocytes with very high efficiency, and demonstrated, for the first time, to be functional. These hepatocytes were characterised at cellular and molecular levels. The GPSC-derived hepatocytes not only expressed hepatic markers, but were also metabolically active as shown by albumin and haptoglobin secretion, urea synthesis, glycogen storage and indocyanine green uptake. Previous studies have revealed some inherent differences in gene expression between undifferentiated mouse ES cells and GPSCs. We wanted to investigate whether this difference may impact on the hepatocyte differentiation capacity of the GPSCs. Large-scale gene expression profiling revealed a strong similarity between GPSC and ES cells at different stages of induced hepatic differentiation. Moreover, Pearson correlation analysis of the microarray datasets revealed that, at late hepatic differentiation stages, the in vitro-derived cells were closer to fetal mouse primary hepatocytes. Thus, adult GPSCs offer great potential for cell ment therapy for a wide variety of liver diseases. Mouse ES cells and GPSCs at various times of hepatocyte differentiation, compared to embryonal (E16) and postnatal (PN1) mouse primary hepatocytes. The supplementary file 'GSE19044_non-normalized.txt' contains non-normalized data for Samples GSM471318-GSM471359.

ORGANISM(S): Mus musculus

SUBMITTER: Sharmila Fagoonee

PROVIDER: E-GEOD-19044 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Hepatocytes obtained from mouse GPSCs are functional and show similar expression profiles to fetal primary hepatocytes

Project description:Germline cell-derived pluripotent stem cells (GPSCs) are similar to embryonic stem (ES) cells in that they can proliferate intensively and differentiate into a variety of cell types, including cardiomyocytes and neurons. In this report, mouse GPSCs were induced to differentiate into hepatocytes with very high efficiency, and demonstrated, for the first time, to be functional. These hepatocytes were characterised at cellular and molecular levels. The GPSC-derived hepatocytes not only expressed hepatic markers, but were also metabolically active as shown by albumin and haptoglobin secretion, urea synthesis, glycogen storage and indocyanine green uptake. Previous studies have revealed some inherent differences in gene expression between undifferentiated mouse ES cells and GPSCs. We wanted to investigate whether this difference may impact on the hepatocyte differentiation capacity of the GPSCs. Large-scale gene expression profiling revealed a strong similarity between GPSC and ES cells at different stages of induced hepatic differentiation. Moreover, Pearson correlation analysis of the microarray datasets revealed that, at late hepatic differentiation stages, the in vitro-derived cells were closer to fetal mouse primary hepatocytes. Thus, adult GPSCs offer great potential for cell ment therapy for a wide variety of liver diseases.

2010-06-01 | GSE19044 | GEO

Gp130-dependent release of acute phase proteins is linked to the activation of innate immune signaling pathways

Project description:Gp130 dependent gene expression was analyzed in a previously established hepatocyte-specific gp130-knockout mouse model. Whole transcriptome analysis for isolated hepatocytes was performed to measure tissue specific responses after proinflammatory stimulus with IL-6 across different time points. We observed differences in the hepatocyte-specific transcriptional gp130 dependent response for genes associated with different aspects of the innate immune system. Our findings suggest a complex network of tightly-linked genes involved in the early activation of different parts of the innate immune response including acute phase proteins, complement and coagulation cascade. Total RNA obtained from a total number of 61 samples of isolated hepatocytes of hepatocyte-specific gp130-knockout and gp130flox mice, which were subjected to Il-6 treatment for 0, 1, 3, 6 or 12 hours, respectively.

2010-11-13 | E-GEOD-25278 | biostudies-arrayexpress

Genome-wide analysis of different response of hepatocytes and oval cells to hepatitis B virus expression during HBV-related hepatocarcinogenesis

Project description:Analysis of genetic changes of hepatocytes and oval cells induced by HBV. The hypothesis tested in the present study was that hepatic progenitor cells (oval cells) are the founder cells of hepatocellular carcinoma (HCC). Results provide important information of the different responses of hepatocytes and oval cells to HBV expression during HBV-related hepatocarcinogenesis, such as up- or down-regulated oncogenes,tumor suppressor genes and signaling pathways.Results also provide new insights into the biology of human HCC. Total RNA obtained from primary cultured hepatocytes and oval cells isolated from the healthy and HBV transgenic mice.

2012-07-11 | E-GEOD-39289 | biostudies-arrayexpress

Identification of myeloid-derived growth factor as a mechanically-induced, growth-promoting angiocrine signal for human hepatocytes

Project description:Recently, we have shown that after partial hepatectomy (PHx), an increased hepatic blood flow initiates liver growth in mice by vasodilation and mechanically-triggered release of angiocrine signals. Here, we use mass spectrometry to identify a mechanically-induced angiocrine signal in human hepatic endothelial cells, that is, myeloid-derived growth factor (MYDGF). We show that it induces proliferation and promotes survival of primary human hepatocytes derived from different donors in two-dimensional cell culture, via activation of mitogen-activated protein kinase (MAPK) and signal transducer and activator of transcription 3 (STAT3). MYDGF also enhances proliferation of human hepatocytes in three-dimensional organoids. In vivo, genetic deletion of MYDGF decreases hepatocyte proliferation in the regenerating mouse liver after PHx; conversely, adeno-associated viral delivery of MYDGF increases hepatocyte proliferation and MAPK signaling after PHx. We conclude that MYDGF represents a mechanically-induced angiocrine signal and that it triggers growth of, and provides protection to, primary mouse and human hepatocytes

2023-12-28 | PXD033942 | Pride

RNA-seq transcriptional profiling in IL6 induced hepatocytes in vitro and livers after 2/3 partial hepatectomy

Project description:We discover IL6, combined with EGF and HGF, promotes long-term expansion (>30 passages in ~150 days with theoretical expansion of ~1E+35 times) of primary mouse hepatocytes in vitro in simple 2D culture, by converting hepatocytes into induced hepatic progenitor cells (iHPCs), which maintain the capacity of differentiation into hepatocytes. IL6 also supports the establishment of single hepatocyte-derived iHPC clones. The summation of the downstream STAT3, ERK and AKT pathways induces a number of transcription factors which support the rapid growth.

2022-11-19 | GSE215423 | GEO

Gene expression profiles in human liver organoid-derived hepatocytes

Project description:Human liver organoids are expected to be a hepatocyte source for preclinical in vitro studies. Although these organoids show long-term proliferation, their hepatic functions remain low. Here, we propose a novel method for two dimensional (2D)-cultured hepatic differentiation from human liver organoids. When cultured under a 2D condition, the single cells from human liver organoids were seeded on collagen type I-coated plates. Then, optimal conditions for hepatic differentiation were screened using several reagents. We determined the 2D-cultured hepatocyte differentiation method from human liver organoids. Hepatic gene expressions in human liver organoids-derived hepatocytes (Org-HEPs) were greatly increased, compared to those in human liver organoids. The metabolic activities of cytochrome P450 (CYP) 1A2, CYP2C8, CYP2E1 and CYP3A4 were at levels comparable to those in primary human hepatocytes (PHHs). These results suggested that human liver organoids could be differentiated into highly functional hepatocytes in 2D culture. We also treated Org-HEPs and PHHs with hepatotoxic drugs. The cell viability of Org-HEPs was almost the same as that of PHHs, suggesting that Org-HEPs could be used for hepatotoxicity tests. Thus, Org-HEPs will be useful for pharmaceutical research.

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In vitro differentiation of hepatoblasts into hepatocytes

Project description:We differentiated primary mouse E14.5 Dlk1+ hepatoblasts in vitro into hepatocyte-like cells that recapitulated morphological features of hepatocytes. To assess the level of differentiation, we performed RNA-seq analysis and compared gene expression profiles of hepatobalsts, in vitro differentiated hepatocytes and mature hepatocytes isolated from adult mouse livers as a control.

2021-07-26 | GSE176069 | GEO

Conversion of terminally committed mouse hepatocytes to culturable bipotent progenitor cells

Project description:A challenge for advancing approaches to liver regeneration is loss of functional differentiation capacity when hepatocyte progenitors are maintained in culture. Recent lineage-tracing studies have shown that mature hepatocytes (MHs) convert to an immature state during chronic liver injury, and we investigated whether this conversion could be recapitulated in vitro and if such converted cells could represent a source of expandable hepatocytes. We report that a cocktail of small molecules, Y-27632, A-83-01 and CHIR99021, can convert rat and mouse MHs in vitro into proliferative bipotent cells, which we term chemically induced liver progenitors (CLiPs). CLiPs can differentiate into both MHs and biliary epithelial cells that can form functional ductal structures. CLiPs in long-term culture did not lose their proliferative capacity or their hepatic differentiation ability, and rat CLiPs were shown to extensively repopulate chronically injured liver tissue. Thus our study advances the goals of liver regenerative medicine. Transcriptomic analysis for mouse CLiPs at P1 which underwent hepatic induction (Matrix 1). Transcriptomic analysis for mouse CLiPs at P11-12 which underwent hepatic induction (Matrix 2).

2016-11-15 | GSE87579 | GEO

CDK5RAP3 regulates hepatocyte differentiation in human

Project description:CDK5RAP3, an UFL1 adaptor for UFMylation, is crucial for liver development in mouse. However, the function of CDK5RAP3 in human hepatocyte differentiation remains unclear. The stepwise generation of hepatocyte-like cells derived from human embryonic cells simulates human hepatocyte differentiation in vitro. Here, the human embryonic cells were conducted to differentiate into hepatocytes and ScRNA-Seq analysis was performed to elucidate the molecular mechanism of CDK5RAP3 in human liver development. We found CDK5RAP3 was important for human hepatocytes differentiation. The CDK5RAP3 deficiency upregulated ER stress in hepatic-endoderm stage, and impaired ribosome homeostasis in mature hepatocytes.

2024-01-13 | GSE253134 | GEO

Human embryonic stem cells as a tool to study hepatocyte differentiation

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