Cell surface n-glycoproteins on human pluripotent stem cell derived hepatic endoderm
Ontology highlight
ABSTRACT:
INSTRUMENT(S): LTQ Orbitrap Velos
ORGANISM(S): Human
SUBMITTER: Rebekah L. Gundry
PROVIDER: MSV000080215 | MassIVE | Mon Oct 03 09:06:00 BST 2016
REPOSITORIES: MassIVE
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Project description:Detailed knowledge of cell surface proteins for isolating well-defined populations of human pluripotent stem cells (hPSCs) would significantly enhance their characterization and translational potential. Normal H9 human embryonic stem cells and the KB3 human induced pluripotent stem cell lines were analyzed by Cell Surface Capture Technology, and in parallel transcript profiles from five independent samples (i.e., Replicas 1-5 for each) were performed to facilitate protein and transcriptomic comparisons. The study compared gene expression profiles of pluripotent stem cells with Cell Surface Capture technology generated N-glycoprotein surfaceome analyses of the same cell types.
2014-03-12 | E-GEOD-55805 | biostudies-arrayexpress
Project description:Detailed knowledge of cell surface proteins for isolating well-defined populations of human pluripotent stem cells (hPSCs) would significantly enhance their characterization and translational potential. Normal H9 human embryonic stem cells and the KB3 human induced pluripotent stem cell lines were analyzed by Cell Surface Capture Technology, and in parallel transcript profiles from five independent samples (i.e., Replicas 1-5 for each) were performed to facilitate protein and transcriptomic comparisons.
2014-03-12 | GSE55805 | GEO
Project description:The future clinical use of embryonic stem cell (ESC)-based hepatocyte replacement therapy depends on the development of an efficient procedure for differentiation of hepatocyes from ESCs. Here we report that a high density of human embryonic stem cell (ESC)-derived fibroblast-like cells (hESdFs) supported the efficient generation of hepatocyte-like cells (HLCs) with functional and mature hepatic phenotypes from primate ESCs and human induced pluripotent stem cells (iPSCs). Molecular and immunocytochemistry analyses revealed that hESdFs caused a rapid loss of pluripotency and induced a sequential endoderm-to-hepatocyte differentiation in the central area of ESC colonies. Knockdown experiments demonstrated that pluripotent stem cells were directed toward endodermal and hepatic lineages by FGF2 and Activin A secreted from hESdFs. Furthermore, we found that the central region of ESC colonies was essential for the hepatic endoderm-specific differentiation, as its removal caused a complete disruption of endodermal differentiation. In conclusion, we describe a novel in vitro differentiation model, and show that hESdF-secreted factors act in concert with regional features of ESC colonies to induce robust hepatic endoderm differentiation in primate pluripotent stem cells. Total RNA were isolated from ORMES6 ESC, differentiated cells at IVDS2 and 3, and cells in the central foci (IVDS2-C) and peripheral (IVDS2-P) area of ESC colonies at IVDS2. Each condition was repeated twice and used ORMES6 ESC as control.
2010-08-20 | E-GEOD-19964 | biostudies-arrayexpress
Project description:Liver is a central organ for drug and xenobiotics metabolism in human body. Due to the differences in interspecies metabolism, human model system for liver metabolism is necessary. However, the hepatic model systems derived from human pluripotent stem cells for metabolic screening and assays did not recapitulate the most of metabolic capabilities of human liver or primary human hepatocytes to date. In this study, we developed human hepatic endoderm organoids which are expandable and subsequently differentiated human hepatic organoids having metabolic capabilities. To investigate the cellular composition and properties of human hepatic organoids in single cell level, we performed single cell RNA-sequencing.
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Project description:Pluripotent stem cells are being actively studied as a cell source for regenerating damaged liver. For long term survival of engrafting cells in the body, not only do the cells have to execute liverspecific function but also withstand the physical strains and invading pathogens. The cellular innate immune system orchestrated by the interferon (IFN) pathway provides the first line of defense against pathogens. The objective of this study is to assess the innate immune function as well as to systematically profile the IFN-induced genes during hepatic differentiation of pluripotent stem cells. To address this objective, we derived endodermal cells (day 5 postdifferentiation), hepatoblast (day 15) and immature hepatocytes (day 21) from human embryonic stem cells (hESC). Day 5, 15 and 21 cells were stimulated with IFN-α and subjected to IFN pathway analysis. Transcriptome analysis was carried out by RNA sequencing. The results showed that the IFN-α treatment activated STAT-JAK pathway in differentiating cells. Transcriptome analysis indicated stage specific expression of classical and non-classical IFNstimulated genes (ISGs). Subsequent validation confirmed the expression of novel ISGs including RASGRP3, CLMP and TRANK1 by differentiated hepatocytes upon IFN treatment. Hepatitis C virus replication in hESC-derived hepatic cells induced the expression of ISGs – LAMP3, ETV7, RASGRP3, and TRANK1. The hESC-derived hepatic cells contain intact innate system and can recognize invading pathogens. Besides assessing the tissue-specific functions for cell therapy applications, it may also be important to test the innate immune function of engrafting cells to ensure adequate defense against infections and improve graft survival.
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Project description:These are 3 TMT10 samples. The indicated "standard" sub-samples all refer to the same biological sample (obtained as a mixture of all other samples), split and tagged as indicated, to play as a reference for normalization.
TMT10 sample1 contains the following sub-samples:
-TAG 126: Standard.
-TAG 127N: Day 3 conditioned medium of replication-incompetent MEFs cultured on VTN coating in RSeT medium.
-TAG 127C: Day 3 conditioned medium of naive human pluripotent stem cells cultured on a MEF feeder-layer in RSeT medium.
-TAG 128N: Day 3 conditioned medium of naive human pluripotent stem cells cultured on a VTN coating in RSeT medium.
-TAG 128C: Not relevant.
-TAG 129N: Not relevant.
-TAG 129C: Not relevant.
-TAG 130N: Not relevant.
-TAG 130C: RSet medium.
-TAG 131: Standard.
TMT10 sample2 contains the following sub-samples:
-TAG 126: Standard
-TAG 127N: Day 3 conditioned medium of replication-incompetent MEFs cultured on VTN coating in RSeT medium.
-TAG 127C: Day 3 conditioned medium of naive human pluripotent stem cells cultured on a MEF feeder-layer in RSeT medium.
-TAG 128N: Day 3 conditioned medium of naive human pluripotent stem cells cultured on a VTN coating in RSeT medium.
-TAG 128C: Not relevant.
-TAG 129N: Day 2 conditioned medium of naive human pluripotent stem cells cultured on a MEF feeder-layer in RSeT medium.
-TAG 129C: Day 2 conditioned medium of naive human pluripotent stem cells cultured on a VTN coating in RSeT medium.
-TAG 130N: Not relevant.
-TAG 130C: RSet medium.
-TAG 131: Standard.
TMT10 sample3 contains the following sub-samples:
-TAG 126: Standard.
-TAG 127N: Day 3 conditioned medium of replication-incompetent MEFs cultured on VTN coating in RSeT medium.
-TAG 127C: Day 3 conditioned medium of naive human pluripotent stem cells cultured on a MEF feeder-layer in RSeT medium.
-TAG 128N: Day 2 conditioned medium of replication-incompetent MEFs cultured on VTN coating in RSeT medium.
-TAG 128C: Day 2 conditioned medium of replication-incompetent MEFs cultured on VTN coating in RSeT medium.
-TAG 129N: Day 2 conditioned medium of naive human pluripotent stem cells cultured on a MEF feeder-layer in RSeT medium.
-TAG 129C: Day 2 conditioned medium of naive human pluripotent stem cells cultured on a VTN coating in RSeT medium.
-TAG 130N: Not relevant.
-TAG 130C: RSet medium.
-TAG 131: Standard.
Detailed culture and processing methods are described in Cesare et al, under submission.
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Project description:Reprogrammed somatic cells offer a valuable source of pluripotent cells that have the potential to differentiate into many cells types and provide a new tool for regenerative medicine. In the present study we differentiated induced pluripotent stem cells (iPS cells) into hepatic cells. We first showed that mouse iPS cells could from a complete liver in mouse embryo (E14.5) including hepatocytes, endothelial cells, sinusoidal cells and resident macrophages. We then designed a highly efficient hepatocyte differentiation protocol using defined factors on human embryonic stem cells (ES cells). This protocol was found to generate more than 80% albumin expressing cells that show hepatic functions and express most of liver genes as shown by microarray analyses. Similar results were obtained when human iPS cells were induced to differentiate following the same procedure. Experiment Overall Design: Total RNA was harvested from the following sources and used for Affymetrix array analysis following manufacturer defined protocols: Experiment Overall Design: 1) human foreskin fibroblasts, ATCC cell line CRL2097, 3 independent cultures Experiment Overall Design: 2) induced pluripotent stem (iPS) cells derived from CRL2097, 3 independent undifferentiated cultures Experiment Overall Design: 3) induced pluripotent stem (iPS) cells derived from CRL2097, 3 independent cultures harvested at day 20 (d20) of a hepatic differentiation protocol Experiment Overall Design: 4) WAO9 human embryonic stem cells, 3 independent undifferentiated cultures Experiment Overall Design: 5) WAO9 human embryonic stem cells, 3 independent cultures harvested at day 20 (d20) of a hepatic differentiation protocol. <br><br>This experiment was reloaded in November 2010 after additional curation
2009-08-07 | E-GEOD-14897 | biostudies-arrayexpress
Project description:Transcriptional profiling of human iPS-HSCs overexpressing LHX2 compared with control iPS-HSCs, which were cocultured with human induced pluripotent stem cell-derived hepatic progenitor cells (iPS-HPCs).
2019-02-14 | GSE122750 | GEO
Project description:Liver is a central organ for drug and xenobiotics metabolism in human body. Due to the differences in interspecies metabolism, human model system for liver metabolism is necessary. However, the hepatic model systems derived from human pluripotent stem cells for metabolic screening and assays did not recapitulate the most of metabolic capabilities of human liver or primary human hepatocytes to date. In this study, we developed human hepatic endoderm organoids which are expandable and subsequently differentiated human hepatic organoids having metabolic capabilities. To investigate the transcriptome of human hepatic organoids, we performed RNA-sequencing.
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Project description:The future clinical use of embryonic stem cell (ESC)-based hepatocyte replacement therapy depends on the development of an efficient procedure for differentiation of hepatocyes from ESCs. Here we report that a high density of human embryonic stem cell (ESC)-derived fibroblast-like cells (hESdFs) supported the efficient generation of hepatocyte-like cells (HLCs) with functional and mature hepatic phenotypes from primate ESCs and human induced pluripotent stem cells (iPSCs). Molecular and immunocytochemistry analyses revealed that hESdFs caused a rapid loss of pluripotency and induced a sequential endoderm-to-hepatocyte differentiation in the central area of ESC colonies. Knockdown experiments demonstrated that pluripotent stem cells were directed toward endodermal and hepatic lineages by FGF2 and Activin A secreted from hESdFs. Furthermore, we found that the central region of ESC colonies was essential for the hepatic endoderm-specific differentiation, as its removal caused a complete disruption of endodermal differentiation. In conclusion, we describe a novel in vitro differentiation model, and show that hESdF-secreted factors act in concert with regional features of ESC colonies to induce robust hepatic endoderm differentiation in primate pluripotent stem cells.
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