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

ABSTRACT: 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

ORGANISM(S): Homo sapiens

PROVIDER: GSE15283 | GEO | 2013/06/07

SECONDARY ACCESSION(S): PRJNA116511

REPOSITORIES: GEO

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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 Three consecutive passages of both ES4CL1 and MR90C2 cells were grown in standard conditions, FACs sorted cells collected, and total RNA isolated. Microarray was performed on a total of 24 samples, (3 replicates of 4 FACs sorted populations from two independent cell lines).

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

2009-10-01 | GSE13877 | GEO

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

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

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

Transcriptome of ESCs, EpiSCs and reverted ES by MM401 (Mll1 inhibitor) #2

Project description:This study describes the transcriptome profiling of: 1) mouse ES cells in LIF/KSR medium; 2)EpiSCs in bFGF/serum-free (KSR) medium; 3) EpiSCs treated with MM401/LIF KSR at D3 and D6 (P2); 4) rES reverted form EpiSC by MM401/LIF KSR treatment at P6, P30 with or without MM401 .

2016-05-21 | GSE73993 | GEO

GCTM-5 positive and negative cells in pancreatic adenocarcinoma cell lines

Project description:Analysis of GCTM-5 positive and negative cells sorted from CFPAC-1 and SW1990 human pancreatic adenocarcinoma cell line. Results provide insight into molecular characteristics of GCTM-5 positive cells.

2017-07-01 | GSE95176 | GEO

Reversible and irreversible anchorage independent growth in skin cells

Project description:We have investigated the regulation of anchorage-independent growth (AIG) by basic fibroblast growth factor (bFGF) and 12-O-tetradecanoyl phorbol-13-acetate (TPA) in JB6 mouse epidermal cells in the context of wound repair versus carcinogenesis responses. bFGF induces an unusually efficient but reversible AIG response, relative to TPA-induced AIG which is irreversible. Distinct global gene expression profiles are associated with anchorage-independent colonies arising from bFGF-stimulated JB6 cells, relative to colonies arising from fully tumorigenic JB6 cells (RT101), including genes exhibiting reciprocal regulation patterns. Thus, while TPA exposure results in commitment to an irreversible and tumorigenic AIG phenotype, the AIG response to bFGF is reversible with essentially complete restoration of normal cell cycle check point control following removal of bFGF from growth medium. These results are consistent with the physiological role of bFGF in promoting wound healing, and suggest that natural mechanisms exist to reverse transformative cellular phenotypes associated with carcinogenesis. Keywords: cell phenotype comparison

2009-06-01 | GSE13979 | GEO

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

Detecting transcription profile associated to mammosphere medium

Project description:Mammosphere medium (Grange et al. Neoplasia 2008;10(12):1433-43) contains epidermal growth factor (EGF), insulin and basic fibroblast growth factor (bFGF) that may influence transcript expression in a way not necessarily related to the assembly of mammspheres. To identify the set of genes associated mammosphere cell growth medium we analyzed two prototypic situations in triplicate experiments: TUBO cells (a mouse breast cancer cell line derived by Balb-neuT mice) grown in DMEM under adherent conditions (TDA), TUBO cells grown in Mammosphere medium under adherent conditions (TMA).

2013-11-30 | GSE26517 | GEO

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