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

ABSTRACT: 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

ORGANISM(S): Homo sapiens

PROVIDER: GSE13877 | GEO | 2009/10/01

SECONDARY ACCESSION(S): PRJNA110489

REPOSITORIES: GEO

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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

Profiling of FACs sorted subpopulations of iPS cells from cell surface marker profile

Project description:Two independent induced pluripotent stem cell lines: ES4CL1 (derived from foreskin) and MR90C2 (derived from lung Fibroblast) were maintained on inactivated mouse embryonic fibroblast (MEF) feeder cells in the presence of ESC media containing DMEM, 20% knock-out serum replacement with 100ng/mL of bFGF. 7 days after seeding, cells were harvested in TrypLE Express and FACs sorted using antibodies detecting the presence of cell surface antigens GCTM-2 and CD9. 4 independent fractions were isolated after cell sorting: P4 (CD9-Negative, GCTM-2 Negative); P5 (CD9-Low, GCTM-2 Low); P6 (CD9-Medium, GCTM-2 medium) and P7 (CD9-High, GCTM-2 High). Each individual experiment was performed in triplicate. Keywords: cell type comparison

2013-06-07 | GSE15283 | 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

Profiling of FACs sorted subpopulations of iPS cells from cell surface marker profile

2013-06-07 | E-GEOD-15283 | biostudies-arrayexpress

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

2010-03-16 | GSE15257 | GEO

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

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

Discovery of progenitor signatures by time series synexpression analysis during Drosophila cell immortalization [RNA-Seq]

Project description:To characterize the sequence of events associated with RasV12 immortalization of Drosophila embryonic cells, we generated transcriptional time series during cell line establishment, from primary cultures until passage (P) 19. We generated three transcriptional time series from three cell lines (R1, R4 and R5) by sampling the cultures at successive stages, early (P2-4), intermediate (P4-11), and late (P16-19), characterized by different passage times. Time points for the R1 time-series were: P2, P3, P4, P5, P7, P8, P10, P11, P16, P17 and P19; for the R4 time-series: P2, P3, P4, P5, P6, P7, P9, P11, P12, P16, P17 and P19; and for the R5 time-series: P2, P3, P4, P6, P7, P8, P16, P17 and P19

2015-09-24 | E-GEOD-73353 | biostudies-arrayexpress

Discovery of progenitor signatures by time series synexpression analysis during Drosophila cell immortalization [Microarray Expression]

Project description:To characterize the sequence of events associated with RasV12 immortalization of Drosophila embryonic cells, we generated transcriptional time series during cell line establishment, from primary cultures until passage (P) 19. We generated two transcriptional time series from two cell lines (R3 and R7) by sampling the cultures at successive stages, early (P2-4), intermediate (P4-11), and late (P16-19), characterized by different passage times. Time points for the R3 time-series were: P2, P2 (replicate), P5, P6, P7, P8, P16, P17 and P19; for the R7 time-series: P2, P2 (replicate), P3, P4, P7, P8, P16, P17 and P19.

2015-09-23 | E-GEOD-73335 | 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

Expression of four hESC lines in enzymatic and mechanic passaging methods

Project description:hESC have morphologic, genetic and genomic alternatiions when cells cultured in different passaging condition. Here transcriptome of four different hESC lines were compared in two passaging methods.

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