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Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Direct comparison of HES2 ESC overexpressing CD30 variant with HES2 overexpressing GFP

ABSTRACT: HES2 ESC were infected with a lentivirus construct containing either CD30 (TNFRSF8) extracellular deficient variant or GFP, using the ViraPowerâ?¢ Promoterless Lentiviral GatewayÂ® Kits (Invitrogen). HES2 cells were recovered and grown for 19 passages. HES2 were commonly grown on Mouse Embryonic Feeder cells in the presence of KSOR media. Before collection of RNA, ES cells were FACs sorted with antibodies to GCMT-2 and CD9 to separate ES cells from differentiated cells and mouse feeders. The experiment was repeated in triplicate. The final aim was to deduce the effect of overexpressing the variant form of CD30 on gene expression in HES2 cells. Three consecutive passages of HES2 cells over-expressing either CD30V or GFP were grown and subject to FACs sorting, collection and microarray. A total of 6 samples, (3 replicates of these 2 populations) were used in the experiment.

ORGANISM(S): Homo sapiens

SUBMITTER: Gabriel Kolle

PROVIDER: E-GEOD-13261 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Similar Datasets

Direct comparison of HES2 ESC overexpressing CD30 variant with HES2 overexpressing GFP

Project description:HES2 ESC were infected with a lentivirus construct containing either CD30 (TNFRSF8) extracellular deficient variant or GFP, using the ViraPower™ Promoterless Lentiviral Gateway® Kits (Invitrogen). HES2 cells were recovered and grown for 19 passages. HES2 were commonly grown on Mouse Embryonic Feeder cells in the presence of KSOR media. Before collection of RNA, ES cells were FACs sorted with antibodies to GCMT-2 and CD9 to separate ES cells from differentiated cells and mouse feeders. The experiment was repeated in triplicate. The final aim was to deduce the effect of overexpressing the variant form of CD30 on gene expression in HES2 cells.

2009-10-10 | GSE13261 | GEO

HES2 embryonic stem cells (ESCs) grown in standard conditions and FACs sorted for GCTM-2 and CD9

Project description:HES2 ESCs were grown in standard ES culture conditions. After 1 week, these cells were FACs sorted for the presence of GCTM-2 and CD9. Keywords: Sorted cell comparison Three consecutive passages of HES2 cells were grown and subjected to FACs sorting. For each, 4 sorted populations of cells (p4, p5, p6, p7) were collected and these were subjected to microarray. A total of 12 samples: 3 replicates each of p4, p5, p6, and p7.

2010-04-08 | E-GEOD-13201 | biostudies-arrayexpress

Membrane-polysome associated fractionation of HES2 embryonic stem cells

Project description:The membrane fraction and the cytosolic fraction of HES2 cells were collected and subjected to microarray. The experiment was performed in triplicate Keywords: membrane-polysome fractionation of RNA Three consecutive passages of HES2 cells were grown and subject to membrane-polysome fractionation. A total of 6 samples, (3 replicates of 2 fractions) were included in the experiment

2010-04-07 | E-GEOD-14037 | biostudies-arrayexpress

Ascorbate effect on serum free cultured hESC

Project description:Ascorbate activates CD30 expression and causes widespread specific demethylation of the epigenome of serum free cultured hESC. The genetic and epigenetic integrity of human Embryonic Stem cells (hESCs) is critical to their future applications in research and medicine. hESC cultured in serum free media can accumulate point mutations, aneuploidy and progressive epigenetic changes over prolonged culture in vitro. We have identified ascorbate as the only molecule in the very widely used Knock-out serum replacement medium that is sufficient to induce expression of CD30, a biomarker for aneuploidy in hESCs. In fact, we show that hESC cultured in the presence of ascorbate for 20 passages not only display demethylation of the CD30 locus, but exhibit widespread and remarkably specific and consistent demethylation of 1,847 genes in both HES2 and HES3 cells. The specific ascorbate induced demethylation changes in the hESC epigenome, of which 86% are shared between the two lines, identify a subset of genes in hESC, including CD30, that are sensitive to serum free culture medium induced epigenetic changes. Experiment. HES2 cells were maintained on 20% Fetal calf serum on mouse embryonic fibroblast feeder layers with mechanical dissection. Using HES2 after 99 mechanical passages (P99), hESC were grown for 17-20 (+17 - +20) passages in the presence of 20% Knock-Out Serum Replacement (Invitrogen) either with (+ASC) or without (-ASC) Ascorbate with enzymatic culturing.

2010-06-10 | GSE16590 | GEO

Ascorbate effect on serum free cultured hESC

2010-06-23 | E-GEOD-16590 | biostudies-arrayexpress

ESC (HES2, MEL1 or H9) grown in various conditions and FACs sorted for GCTM-2 and CD9

Project description:In the following experiment, three different hESC cell lines (HES2, MEL1 and H9) were grown in the presence of KOSR, KOSR or mTESR containing media respectively. KOSR (Knockout serum replacement medium) is a standard media allowing the growth of hESC without the need for manual passaging - Enzymatic passaging is used instread. mTESR (Ludwig et al., 2007) is a media allowing the growth of hESC on matrigel with enzymatic passaging. At day 7 after passaging, these cells were FACs sorted for the presence of GCTM-2 and CD9 into 4 distinct fractions (p4: GCTM-2-neg, CD9-neg; p5: GCTM-2-low, CD9-low; p6: GCTM-2-medium, CD9-medium and p7: GCTM-2-high, CD9-high). For each cell line-subfraction combination, RNA was harvested and subject to microarray. From each experiment (individual cell line), 4 samples were collected (p4, p5, p6 and p7) and these were subject to microarray. In each case the experiment was performed in triplicate Three independent experiments of three consecutive passages of ESC cells were grown and subject to FACs sorting, collection and microarray. A total of 36 samples, (3 experiments of 3 replicates of 4 sorted populations of cells (p4, p5, p6, p7))

2010-04-07 | E-GEOD-13877 | biostudies-arrayexpress

Homo sapiens

Project description:Direct comparison of HES2 ESC overexpressing CD30 variant with HES2 overexpressing GFP

| PRJNA109549 | ENA

Isolation and Characterisation of Renal Precursor Cells Derived from Human Embryonic Stem Cells

Project description:HES4 hESC were cultured in serum media and maintained on a layer of mouse embryonic fibroblast feeder cells at a density of 6 x 104 cells/cm2. For differentiation: hESC were differentiated for a total of 14 days. Differentiation was induced by passaging 4 human ES cell pieces onto 12 well plates seeded with 0.67 x 10E4 cells/cm2. Cells were maintained in media containing 20% FCS for 2 days before media containing 5% FCS was used. Reduced serum media was changed every second day for the remaining 12 days ESC cells were taken at time zero and RNA was isolated (hESC_undiff). Cells differentiated by the above procedure at day 14 were isolated and RNA extracted (Diff). The remaining cells were FACs sorted with antibodies to Podocalyxin, CD24 and GCTM-2. Two populations were isolated, 1: Positive for podocalyxin and CD24 and low level of expression of GCTM-2 (POD+CD24+GCTM2Low) and 2: Positive for podocalyxin and CD24 and negative for GCTM-2 (POD+CD24+GCTM2Neg). The experiment was repeated in triplicate. The final aim was to determine the gene expression enrichment in cells with the marker profile (Podocalyxin+, CD24+, GCTM-2-neg) which is predicted to be enriched for kidney precursors. Keywords: cell type comparison Three consecutive passages of HES4 cells were grown in standard conditions, RNA derived or differentiated for 14 days subject to FACs sorting, collection and RNA isolated. Microarray was performed on a total of 12 samples, (3 replicates of 4 populations).

2010-03-16 | E-GEOD-15257 | biostudies-arrayexpress

HES2 embryonic stem cells (ESCs) grown in standard conditions and FACs sorted for GCTM-2 and CD9

Project description:HES2 ESCs were grown in standard ES culture conditions. After 1 week, these cells were FACs sorted for the presence of GCTM-2 and CD9. Keywords: Sorted cell comparison

2009-10-01 | GSE13201 | GEO

Embryonic stem cells culture conditions support distinct ES cell states associated with specific developmental stages and cell types

Project description:Identification of transcripts in murine embryonic stem (ES) cell lines growing under 3 different self-renewing conditions; GMEM + 10% FCS, N2B27 + 2i (1 μM PD032 and 3 μM CHIR99021) and knockout serum replacement (KOSR) medium (80% Knockout DMEM, 20% Knockout Serum Replacement). Under all conditions, ES cells were grown on gelatin-coated dishes and passaged by trypsinisation. ES cells were cultured in each condition for at least 3 passages prior to sample collection. The aim of this array experiment is to identify significant differences in transcript levels of ES cells grown under different conditions. Differences of transcript levels from the different conditions should be consistent among the biological and technical replicates for each.

2016-07-14 | GSE72445 | GEO

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