Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of primary human tubular epithelial cells (HUTEC) response to TGFbeta at three concentrations to study the epithelial to mesenchymal transition

ABSTRACT: This experiment was aimed to study the Epithelial to mesenchimal transition (EMT) induced by TGFbeta in primary tubular epithelial cells. Primary cultures were grown for 4 days using three different TGFbeta1 concentrations: 5 ng/ml, 10 ng/ml and 50 ng/ml. Cells were grown in plastic flasks (Falcon) or in plastic flasks (Falcon) covered by type I collagen. Differentially expressed genes were identified comparing treated vs. control cells maintained for 4 days in 1% serum without TGFbeta1.

INSTRUMENT(S): ScanArray Lite [PerkinElmer]

ORGANISM(S): Homo sapiens

SUBMITTER: Stefano Campanaro

PROVIDER: E-MEXP-566 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Understanding Natural Killer (NK) cell anatomical distribution is key to dissect the role of these unconventional lymphocytes in physiological and disease conditions. In mouse, NK cells have been detected in various lymphoid and non-lymphoid organs, while in humans the current knowledge of NK cell distribution at steady state is mainly restricted to lymphoid tissues. The translation to humans of findings obtained in mice is facilitated by the identification of NK cell markers conserved between these two species. The Natural Cytotoxicity Receptor (NCR) NKp46 is a marker of the NK cell lineage evolutionary conserved in mammals. In mice, NKp46 is also present on rare T cell subsets and on a subset of gut Innate Lymphoid Cells (ILCs) expressing the retinoic acid receptor-related orphan receptor gammat (RORgammat) transcription factor. Here, we documented the distribution and the phenotype of human NKp46+ cells in lymphoid and non-lymphoid tissues isolated from healthy donors. Human NKp46+ cells were found in splenic red pulp, in lymph nodes, in lungs and gut lamina propria, thus mirroring mouse NKp46+ cell distribution. We identified a novel cell subset of CD56dimNKp46low cells that includes RORgammat+ILCs with a lineage-CD94-CD117brightCD127bright phenotype.We also included data regarding the genome-wide transcriptional profiles of human healthy colonic NK cells and RORgammat+ILCs.The use of NKp46 thus contributes to establish the basis for analyzing quantitative and qualitative changes of NK cell and ILC subsets in human diseases. Human colonic CD56-NKp46lowCD117brightCD127bright cells = LTi cells and CD56+NKp46+CD117-CD127- cells =NK cells were isolated from macroscopically unaffected areas of colon of patients with colon cancer. Indicated populations were sorted and immediately lysed in RLT buffer supplemented with 10% beta-mercaptoethanol (Quiagen, France). Lysates of three distinct donors were pooled and RNA was isolated by using RNAeasy Microkit (Quiagen, France). Duplicates were performed for each cell type. cRNA were obtained after double amplification using the MessageAmp II aRNA Amplification Kit (Ambion, France). cRNA were then hybridized on Human Genome HG_U133 +2.0 Affymetrix chips. Chip images were generated using Affymetrix AGCC 3.2 software, then expression data were extracted and normalized using Affymetrix Expression console 1.1 with the algorithm RMA. Data obtained were expressed as log2.

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Transcription profiling of primary human tubular epithelial cells (HUTEC) response to TGFbeta at three concentrations to study the epithelial to mesenchymal transition

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