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Genome-wide analysis of the gene expression profile of in vivo induced pluripotent stem cells

ABSTRACT: Analysis of the gene expression profile of a novel type of pluripotent stem cells (in vivo induced pluripotent stem cells, named i2PS cells) obtained by reprogramming BALB/c mice hepatocytes in vivo. The hypothesis tested in the present study is that in vivo induced pluripotent stem cells will have a similar gene expression profile to that of other pluripotent stem cells. Hence the embryonic stem cell line E14TG2A is used as a reference in this study. Results provide valuable information for the overall molecular characterisation of cells that have been reprogrammed to pluripotency in vivo, together with the confirmation of their pluripotent character at the gene expression level. Total RNA was obtained from i2PS and E14TG2A cells cultured under standard embryonic stem cells conditions (i.e. DMEM supplemented with LIF). Three biological replicates were included for each cell type. Data were analysed using the gene expression levels of E14TG2A cells as a reference.

ORGANISM(S): Mus musculus

SUBMITTER: Kostas Kostarelos

PROVIDER: E-GEOD-55996 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Genome-wide analysis of the gene expression profile of in vivo induced pluripotent stem cells

Project description:Analysis of the gene expression profile of a novel type of pluripotent stem cells (in vivo induced pluripotent stem cells, named i2PS cells) obtained by reprogramming BALB/c mice hepatocytes in vivo. The hypothesis tested in the present study is that in vivo induced pluripotent stem cells will have a similar gene expression profile to that of other pluripotent stem cells. Hence the embryonic stem cell line E14TG2A is used as a reference in this study. Results provide valuable information for the overall molecular characterisation of cells that have been reprogrammed to pluripotency in vivo, together with the confirmation of their pluripotent character at the gene expression level.

2014-03-19 | GSE55996 | GEO

Comparison of hPSCs-derived cholangiocyte-like cells with primary biliary tissue.

Project description:We established a step-wise protocol for the generation of human induced pluripotent stem cells (hPSCs) derived cholangiocyte-like cells (CLCs). The aim of this experiment was to provide in depth characterisation of the transcriptome of CLCs. These cells were compared with two of their in vitro progenitors, hPSCs and hepatoblasts (HBs), and to freshly isolated primary biliary tissue.

2015-05-26 | E-MTAB-2965 | biostudies-arrayexpress

Metabolomics Workbench ST000003 - GNPS - Metabolomic analysis of mouse embryonic fibroblasts, embryonic stem cells, and induced pluripotent stem cells

Project description:mouse embryonic fibroblasts, embryonic stem cells, and induced pluripotent stem cells were compared via metabolomic analysis

2018-08-02 | MSV000082680 | GNPS

HipSci project proteomics batch 1, comprising 6 IPS cell lines from 3 donors

Project description:The Human Induced Pluripotent Stem Cells Initiative (HipSci) is generating a large, high-quality reference panel of human IPSC lines. This is a submission of mass-spectrometry analyses from 6 induced pluripotent stem cell lines generated by the HipSci project.

2017-05-15 | MSV000081088 | MassIVE

The subcellular proteome of mouse pluripotent embryonic stem cells

Project description:Our understanding of the biology of embryonic stem (ES) cells is deeply rooted in characterization of their transcriptomes, epigenetics and underlying gene regulatory networks. There is evidence that post-transcriptional processes such as signaling, adhesion, protein turnover and post translational modifications make a significant contribution to regulating the balance between self-renewal and differentiation, and it is therefore necessary to also characterize ES cells at the protein level. In this experiment, we used a workflow termed hyperLOPIT (hyperplexed localization of organelle proteins by isotope tagging) to characterize the subcellular distribution of proteins in a population of self-renewing E14TG2a mouse ES cells. Over 5,000 protein groups were quantified in both of the two replicates, enabling characterization of protein localization to organelles (including sub-nuclear resolution), cell surface, cytoskeleton and cytosol. The steady-state localization of transitory proteins, protein complex constituents, and signaling cascades could also be mapped.

2016-01-14 | PXD001279 | Pride

Human pluripotent stem cell-derived ROR1e lens epithelial cell single cell RNA-sequencing

Project description:Detailed transcriptomic analyses of differentiated cell populations derived from human pluripotent stem cells is routinely used to assess the identity and utility of the differentiated cells. In particular, single cell RNA-sequencing (scRNA-seq) can provide insights into both the cellular and transcriptional heterogeneity of differentiated cell populations. Here we provide scRNA-seq data obtained from ROR1-expressing lens epithelial cells (ROR1e LECs) obtained via directed differentiation of CA1 human embryonic stem cells. Through the use of principal component analysis, heat maps and gene ontology assessments, we demonstrate that ROR1e LECs represent a highly purified and large-scale population of lens cells. These data provide a resource for future characterisation of both normal and cataractous human lens biology.

2020-11-27 | E-MTAB-9178 | biostudies-arrayexpress

HipSci project proteomics batch 1, comprising 6 IPS cell lines from 3 donors

2017-05-12 | PXD005506 | Pride

Wnt signalling sustains an EpiSCs subpopulation similar to primitive streak with increased mesendodermal potency

Project description:During gastrulation, epiblast cells are pluripotent and their fate is thought to be constrained principally by their position. Cell fate is progressively restricted by localised signalling cues from areas including the primitive streak (PS). However, it is unknown whether this restriction accompanies, at the single cell level, a reduction in potency. Investigation of these early transition events in vitro is possible via the use of Epiblast Stem Cells (EpiSCs), self-renewing pluripotent cell lines equivalent to the postimplantation epiblast. Strikingly, EpiSCs express various early lineage-specific markers in self-renewing conditions. However, it is unknown whether cells that express these markers are pluripotent, spontaneously differentiated, or biased towards specific lineages. Here we show that EpiSC are inherently heterogeneous and contain two major and mutually exclusive subpopulations with PS and neural characteristics respectively. Using differentiation assays and embryo grafting we demonstrate that PS-like EpiSCs are biased towards mesoderm and endoderm differentiation but they still retain their pluripotent character. The acquisition of a PS character by undifferentiated EpiSC is mediated by paracrine Wnt signalling. Elevation of Wnt activity promotes further restriction into PS-associated cell fates which occurs via the generation of distinct clonal mesendodermal and neuromesodermal precursors. Collectively, our data suggest that primed pluripotency encompasses a range of reversible lineage-biased states reflecting the birth of pioneer lineage precursors from a pool of uncommitted EpiSCs similar to the earliest cell fate restriction events taking place in the gastrula-stage epiblast. Total RNA obtained (3 biological replicates) from flow sorted dsRed2+, dsRed2-, and +CHIRON treated for 48h cells was isolated and labelled/amplified using the IlluminaM-BM-. TotalPrepM-bM-^DM-" RNA Amplification Kit (Life Technologies)

2014-03-04 | E-GEOD-48476 | biostudies-arrayexpress

Defining Essential Regulators of Naïve Human Pluripotent Cell Reprogramming

Project description:Defining how human pluripotent cell identity is controlled and in particular how naïve pluripotency is acquired during cell reprogramming is crucial for the future applications of pluripotent stem cells. However, the regulatory pathways of naïve cell reprogramming remain incompletely understood. Here, we used genome-wide CRISPR-Cas9 screening to identify novel regulators of primed to naïve pluripotent stem cell reprogramming, including genes that are essential for reprogramming and genes that normally impede reprogramming and whose targeted deletion led to enhanced reprogramming. Integrated analysis defined specific chromatin complexes and signalling pathways as critical regulators of naïve reprogramming, and were largely distinct from regulators of somatic cell reprogramming. Mechanistically, PRC1.3 and PRDM14 are jointly required to transcriptionally repress developmental and gene regulatory factors to ensure naïve cell reprogramming. Additionally, small molecule inhibitors of reprogramming impediments increased the efficiency of naïve cell reprogramming, and are of practical benefit that can improve on current reprogramming methods. Taken together, we have identified novel regulators controlling the establishment of naïve pluripotency in human cells, which will open up new ways to exploit the full potential of pluripotent stem cells. These results also provide new insights into mechanisms that destabilise and reconfigure cell identity during cell state transitions.

2022-04-06 | PXD023547 | Pride

Transcriptome data from induced Pluripotent Stem cells

Project description:We have generated human induced Pluripotent Stem cells (hiPSc) using Sendai virus-mediated delivery of reprogramming factors. hiPSc lines have been screened using SNP array to assess chromosomal stability (alongside the fibroblast lines from which they derived), and validation of the pluripotency of the hiPSc lines is provided by Pluritest assessment of transcriptome datasets, prior to differentiation and downstream assays. Single cell RT-qPCR analysis of induced pluripotent stem cell-derived cortical neurons reveals frequent dual layer identity. Handel AE, Chintawar S, Lalic T, Whiteley E, Giustacchini A, Argoud K, Sopp P, Bowden R, Cowley SA, Newey S, Ackerman C, Ponting CP and Cader ZM. Submitted human iPSc lines were derived from human dermal fibroblasts from control donors. SNP and gene expression datasets from control lines. Note that iPS NHDF-1, also used in the current study, has been published previously [Van Wilgenburg B., Browne C., Vowles J. & Cowley S. A. Efficient, long term production of monocyte-derived macrophages from human pluripotent stem cells under partly-defined and fully-defined conditions. PLoS One 8, e71098 (2013); Physiological characterisation of human iPS-derived dopaminergic neurons. Hartfield EM, Yamasaki-Mann M, Ribeiro Fernandes HJ, Vowles J, James WS, Cowley SA, Wade-Martins R. PLoS One. 2014 9(2):e87388]

2015-05-28 | E-GEOD-69288 | biostudies-arrayexpress

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