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Undifferentiated mouse embryonic stem cells, differentiated nestin-positive cells and fibroblast feeder layer.

ABSTRACT: Embryonic stem (ES) cells and ES cell-derived progeny characterized by nestin expression (including neural progenitors) were studied (three independent experiments). The mouse ES cell line R1 was cultured on a feeder layer of mouse embryonic fibroblasts (FL). ES cells were differentiated into nestin-positive cells for 4+8 days and 4+11 days according to the differentiation protocol by Rolletschek et al. (Mechanisms of Development 105, 93-104, 2001). The study was performed with three independent cell cultures (= triplicates). RNA was prepared from both undifferentiated ES cells and ES cell-derived nestin-positive cells, and from fibroblast feeder cells. Keywords = ES cell Keywords = nestin positive cells Keywords = feeder layer Keywords: other

ORGANISM(S): Mus musculus

PROVIDER: GSE2375 | GEO | 2005/09/17

SECONDARY ACCESSION(S): PRJNA91531

REPOSITORIES: GEO

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Transcription profiling of mouse undifferentiated embryonic stem cells, differentiated nestin-positive cells and fibroblast feeder layer.

Project description:Embryonic stem (ES) cells and ES cell-derived progeny characterized by nestin expression (including neural progenitors) were studied (three independent experiments). The mouse ES cell line R1 was cultured on a feeder layer of mouse embryonic fibroblasts (FL). ES cells were differentiated into nestin-positive cells for 4+8 days and 4+11 days according to the differentiation protocol by Rolletschek et al. (Mechanisms of Development 105, 93-104, 2001). The study was performed with three independent cell cultures (= triplicates). RNA was prepared from both undifferentiated ES cells and ES cell-derived nestin-positive cells, and from fibroblast feeder cells.

2008-06-12 | E-GEOD-2375 | biostudies-arrayexpress

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Project description:Undifferentiated mouse embryonic stem cells, differentiated nestin-positive cells and fibroblast feeder layer.

| PRJNA91531 | ENA

Gene Expression Analysis of Neural Stem Cells

Project description:Tissue from the telencephalon was isolated from E13.5 BALB/C mouse and allowed to culture as neurospheres in the presence of FGF2. These cultures were assessed for undifferentiated neural stem cells by the expression of Nestin and were found to be ~98% Nestin positive. Comparisons of these nestin positive neural stem cells will be made to R1 ES cells to identify the genes that are important in totipotent, self-renewing ES cells vs. commitment to the multipotent, self-renewing neural stem cell phenotype. Keywords: other

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Comparison between Nestin+ and Nestin- Ptch1 deficient GNPs

Project description:The intermediate filament protein Nestin serves as a biomarker for stem cells and has been used to identify subsets of cancer stem-like cells. However, the mechanistic contributions of Nestin to cancer pathogenesis are not understood. Here we report that Nestin binds the hedgehog pathway transcription factor Gli3 to mediate the development of medulloblastomas of the hedgehog subtype. In a mouse model system, Nestin levels increased progressively during medulloblastoma formation resulting in enhanced tumor growth. Conversely, loss of Nestin dramatically inhibited proliferation and promoted differentiation. Mechanistic investigations revealed that the tumor-promoting effects of Nestin were mediated by binding to Gli3, a zinc finger transcription factor that negatively regulates hedgehog signaling. Nestin binding to Gli3 blocked Gli3 phosphorylation and its subsequent proteolytic processing, thereby abrogating its ability to negatively regulate the hedgehog pathway. Our findings show how Nestin drives hedgehog pathway-driven cancers and uncover in Gli3 a therapeutic target to treat these malignancies. Nestin+ and Nestin- GNPs (granule neuron precursors) were purified from Nestin-CFP/Math1-Cre/Ptch1-loxp cerebella at postnatal day 4 by FACs, and total RNA from these two cell populations were extracted, and then labeled and hybridized to Affymetrix Mouse Genome 430 2.0 arrays.

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Project description:A monolayer of human nestin positive cells grown in culture using bFGF and EGF was compared to hand picked isolated islets.

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Comparison between Nestin+ and Nestin- Ptch1 deficient GNPs

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Gene expression of human corneal epithelial cells grown with irradiated human fibroblasts or irradiated mouse 3T3 feeder layer .

Project description:human corneal epithelial cells were isolated from healthy human donor eyes. Cells were cultured with irradiated 3T3 (i3T3) murine fibroblast feeder cells, irradiated human corneal fibroblasts (iHFL) or without feeder layer

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Methanol fixed feeder layers altered the pluripotency and metabolism of bovine pluripotent stem cells

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Human iPSCs derived under feeder free conditions displays an unqiue cell cycle and DNA replication genotype

Project description:Induced pluripotent stem cells (iPSCs) have been generated from various somatic cells under feeder-layer conditions. These feeder-derived iPSCs generated in different labs exhibit greater variability than between different traditional embryo derived hESC lines. For that reason, it is important to develop a standard and defined system for deriving autologous patient stem cells. We have generated iPSCs under feeder-free conditions using Matrigel coated vessels in chemically defined medium, mTeSR1. These feeder-free derived iPSCs are in many ways similar to feeder-derived iPSCs and also to hESCs, with respect to their pluripotent gene expression (OCT4, NANOG, SOX2), protein expression (OCT4, NANOG, SSEA4, TRA160) and differentiation capabilities. We conducted a whole genomic transcript analysis using Affymetrix Human Gene 1.0 ST arrays to elucidate the important differences between traditional feeder-derived iPSCs and feeder-free derived iPSCs. We reveal that feeder-free iPSCs have over-represented terms belonging to DNA replication and cell cycle genes which are lacking in feeder-derived iPSCs. Feeder-free iPSCs are in many aspects more similar to hESCs including; apoptosis, chromatin modification enzymes and mitochondrial energy metabolism. We have also identified potential biomarkers for fully reprogrammed iPSCs (FRZB) and partially reprogrammed iPSCs (POTEG, MX2) based on their expression trends across all cell types. In conclusion, feeder-free derived iPSCs is transcriptomically more similar to hESCs than feeder derived iPSCs, in many biological functions.

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