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Embryonic Stem Cell Differentiation LIF vs no LIF

ABSTRACT: Description: The pluripotency of an embryonic stem cell makes it an attractive target for differentiation strategies aimed at producing tissues for therapeutic purposes. Yet, even though we know that ES cells have tremendous capacity for differentiation, we do not fully understand the molecular paths taken by these cells as they differentiate. In addition, it is unclear how important the removal of leukemia inhibitory factor is to ES cell differentiation. Moreover, it is unclear how the path of ES cell differentiation relates to the molecular decisions made by tissues undergoing normal development. The motivation behind this work is to increase our understanding of this relationship and its potential limitations. Dataset is a murine embryonic stem cell differentiation timecourse consisting of 14 timepoints spanning 17 days of differentiation. Samples were harvested at 24 hour intervals on days 0,1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 13, 15, and 17. There two replicates of each sample in this series. Additionally, there are 14 identically stated differentiation samples, where growth was conducted in ES cell media containing LIF. A common reference design was employed. Groups of assays that are related as part of a time series. Compound Based Treatment: Treatment or lack of treatment with LIF Elapsed Time: Samples were harvested at 24 hour intervals on days 0,1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 13, 15, and 17 Keywords: time_series_design

ORGANISM(S): Mus musculus

PROVIDER: GSE9048 | GEO | 2010/09/09

SECONDARY ACCESSION(S): PRJNA102557

REPOSITORIES: GEO

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Embryonic Stem Cell Differentiation LIF vs no LIF

Project description:Description: The pluripotency of an embryonic stem cell makes it an attractive target for differentiation strategies aimed at producing tissues for therapeutic purposes. Yet, even though we know that ES cells have tremendous capacity for differentiation, we do not fully understand the molecular paths taken by these cells as they differentiate. In addition, it is unclear how important the removal of leukemia inhibitory factor is to ES cell differentiation. Moreover, it is unclear how the path of ES cell differentiation relates to the molecular decisions made by tissues undergoing normal development. The motivation behind this work is to increase our understanding of this relationship and its potential limitations. Dataset is a murine embryonic stem cell differentiation timecourse consisting of 14 timepoints spanning 17 days of differentiation. Samples were harvested at 24 hour intervals on days 0,1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 13, 15, and 17. There two replicates of each sample in this series. Additionally, there are 14 identically stated differentiation samples, where growth was conducted in ES cell media containing LIF. A common reference design was employed. Groups of assays that are related as part of a time series. Compound Based Treatment: Treatment or lack of treatment with LIF Elapsed Time: Samples were harvested at 24 hour intervals on days 0,1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 13, 15, and 17 Keywords: time_series_design Using regression correlation

2010-09-09 | E-GEOD-9048 | biostudies-arrayexpress

Genome-wide analysis of Embryonic Stem Cell Differentiation

Project description:Description: The pluripotency of an embryonic stem cell makes it an attractive target for differentiation strategies aimed at producing tissues for therapeutic purposes. Yet, even though we know that ES cells have tremendous capacity for differentiation, we do not fully understand the molecular paths taken by these cells as they differentiate. Moreover, it is unclear how the path of ES cell differentiation relates to the molecular decisions made by tissues undergoing normal development. The motivation behind this work is to increase our understanding of this relationship and its potential limitations. Overall Design: Dataset is a murine embryonic stem cell differentiation timecourse consisting of 14 timepoints spanning 17 days of differentiation. Samples were harvested at 24 hour intervals on days 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 13, 15, and 17. There two replicates of each sample. A common reference design was employed. Groups of assays that are related as part of a time series. Elapsed Time: amples were harvested on days 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 13, 15, and 17 Keywords: time_series_design

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Genome-wide analysis of Embryonic Stem Cell Differentiation

Project description:Description: The pluripotency of an embryonic stem cell makes it an attractive target for differentiation strategies aimed at producing tissues for therapeutic purposes. Yet, even though we know that ES cells have tremendous capacity for differentiation, we do not fully understand the molecular paths taken by these cells as they differentiate. Moreover, it is unclear how the path of ES cell differentiation relates to the molecular decisions made by tissues undergoing normal development. The motivation behind this work is to increase our understanding of this relationship and its potential limitations. Overall Design: Dataset is a murine embryonic stem cell differentiation timecourse consisting of 14 timepoints spanning 17 days of differentiation. Samples were harvested at 24 hour intervals on days 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 13, 15, and 17. There two replicates of each sample. A common reference design was employed. Groups of assays that are related as part of a time series. Elapsed Time: amples were harvested on days 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 11, 13, 15, and 17 Keywords: time_series_design Using regression correlation

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CEA_SGF:E00005#LIF

Project description:In order to identify the genes involved in LIF/gp130 self-renewal response, we analyzed the transcriptome of Gs2 ES cells induced to differentiate upon LIF withdrawal for 16h, 24h and 48h (LIF16, LIF24 and LIF48 respectively)and compared them to that of undifferentiated ES Gs2 cells continuously maintained in the presence of LIF. Since this cell line also carry a tetracycline regulatable dominant negative form of Stat3 (Stat3F, all the experiment was done in the presence of Tetracycline (tet ON) to avoid Stat3F expression. Keywords: time-course

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RNA-seq analysis of ES cell populations with different functional potentials in serum/LIF and 2i/LIF, as well as analysis of the transcriptional effect of LIF in ground state (2i) culture conditions.

Project description:We report that ES cells cultured in ground state (2i and 2i/LIF) culture conditions are heterogeneous and show heterogeneus expression of extraembryonic markers. Using a highly sensitive reporter for the endoderm marker Hex we can sort Hex high and low populations from either serum/LIF or 2i/LIF and demonstrate that they have different functional properties. Here we explored the transcriptional basis of these functional differences and noted that Hex low (HV-) and Hex high (HV+) populations showed more distinct expression profiles in 2i/LIF than in serum/LIF. Additionally in 2i/LIF the HV+ population showed an upregulation of extraembryonic markers (such as trophoblast stem cell specific genes) and also imprinted genes compared to the HV- population, which is not observed when these populations are sorted from serum/LIF. We also analysed the transcriptional effect of LIF in 2i by analysing unsorted ES cells cultured in either 2i alone or 2i with LIF. We observed that the addition of LIF led to an upregulation of extraembryonic markers but did not effect the expression of pluripotency genes, other than Klf4. Additionally, the most significantly upregulated genes from 2i/LIF cultured ES cells compared to 2i cultured ES cells showed the greatest correlation to placental tissue when compared to the GNF tissue specific expression database. This analysis, alongside functional experiments, suggested that HV+ ES cells in 2i/LIF corresponded to an extraembryonically primed population of cells and that the addition of LIF supported this population.

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Hippocampal circadian proteomes of young and middle-aged mice

Project description:Hippocampal tissues from young and middle-aged C57BL/6J mice were harvested at 4-hour intervals over two days and processed for proteomic analysis using label-free quantification.

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Wnt & LIF signalling in mESC

Project description:The molecular processes underlying the properties of ESC are yet unknown even when it's well established that LIF/STAT3 is neccesary for the maintenance of pluripotency. Other pathways as Wnt are may be implicated in the regulation of the biological mechanisms in mESC. Work model: D3-ES cultivated with or without LIF and treated with chronic (7 days) low doses (50nM) of GSK3 β inhibitor (lithium). We used microarrays to detail the global profile of gene expression to identified different groups of up and down-regulated genes in the work model.

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Effect of LIF on ES cells overexpressing Akt

Project description:Embryonic stem (ES) cells can self-renew indefinitely without losing their differentiation ability to any cell types. Phosphoinositide-3 kinase (PI3K)/Akt signaling plays a pivotal role in various stem cell systems, including the formation of embryonic germ (EG) cells from primordial germ cells and self-renewal of neural stem cells. Here, we show that myristoylated, active form of Akt (myr-Akt) maintained the undifferentiated phenotypes in mouse ES cells without the addition of leukemia inhibitory factor (LIF). The effects of myr-Akt were reversible, because LIF dependence and pluripotent differentiation activity were restored by the deletion of myr-Akt. In addition, myr-Akt-Mer fusion protein, whose enzymatic activity is controlled by 4-hydroxy-tamoxifen, also maintained the pluripotency of not only mouse but also cynomolgus monkey ES cells. These results clearly demonstrate that Akt signaling sufficiently maintains pluripotency in mouse and primate ES cells, and support the notion that PI3K/Akt signaling axis regulates 'stemness' in a broad spectrum of stem cell systems. Keywords: other

2005-04-12 | GSE2516 | GEO

Transcription profiling of mouse embryonic stem cell line CGR8 treated with Leukemia Inhibitory Factor (LIF) to identify LIF-dependent genes and early differentiation markers

Project description:This experiment is part of the FunGenES project (FunGenES - Functional Genomics in Embryonic Stem Cells partially funded by the 6th Framework Programme of the European Union, http://www.fungenes.org). The experiment was conducted at the University Bordeaux 2 (CNRS UMR 5164). The purpose of the experiment was to identify the LIF-dependent genes and early differentiation markers in mouse ES cells. Leukemia Inhibitory Factor (LIF) is a member of the Interleukin-6 (IL-6) family of cytokines, which displays pleiotropic functions, depending on both cell maturity and cell type. LIF maintains ES cell pluripotency and is a critical cell survival factor in myocytes. Materials and methods: CGR8 cell line was used.6 cell conditions tested: + LIF, reinduced 25 min with serum + LIF reinduced 25 min with serum and LIF 500pM - LIF for 24h, reinduced 25 min with serum - LIF for 24h, reinduced 25 min with serum and LIF 500pM- LIF for 48h, reinduced 25 min with serum - LIF for 48h, reinduced 25 min with serum and LIF 500pM. Relationships between samples. 5 independent preparations of mRNA have been done on five different CGR8 passages. Culture conditions: Monolayer adherent cells. RNA isolation method: Qiagen column

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