Zuklys etal_miRNA_C57BL6_TECs
Ontology highlight
ABSTRACT:
ORGANISM(S): Mus musculus
SUBMITTER: Saulius Zuklys
PROVIDER: E-MEXP-3308 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:mTEC and cTEC profiles from 2 week old Dicer fl/fl (WT) and Foxn1Cre::Dicer fl/fl mice. cTEC(CD45-EpCAM+MHCII+UEA-1-Ly51+) and mTEC(CD45-EpCAM+MHCII+UEA-1+Ly51-) were isolated from thymus tissue by cell sorting
2012-09-03 | E-MEXP-3303 | biostudies-arrayexpress
Project description:We compared in vitro mTECs-high from MyD88fl/fl x Foxn1Cre and MyD88fl/fl mice: Thymi were enzymaticaly digested, cells were MACS enriched for CD45- fraction, and FACS sorted using BD Influx sorter. mTECs-high were gated as EpCAM+CD11c-Ly51-MHCII+CD80+. Total RNA was isolated using RNeasy Plus Micro Kit (Qiagen). 5 samples per condition were used.
2020-03-20 | E-MTAB-8024 | biostudies-arrayexpress
Project description:We compared in vitro mTECs-high stimulated with TLR9 ligand CpG ODN (1826) or non stimulated from MyD88 -/- and MyD88 +/+ mice: Thymi were enzymaticaly digested, cells were MACS enriched for CD45- fraction, and FACS sorted using BD Influx sorter. mTECs-high were gated as EpCAM+CD11c-Ly51-MHCII+CD80+. Cell from MyD88 -/- and MyD88 +/+ mice were incubated in RPMI with or without CpG ODN (1826) for 24 hours. Total RNA was isolated using RNeasy Plus Micro Kit (Qiagen). 4 samples per condition were used.
2020-03-20 | E-MTAB-8025 | biostudies-arrayexpress
Project description:We have compared the transcriptional profiles from two epithelial cell types to find genes specifically expressed in thymic medullary epithelial cells. Epithelial cells from thyroid and thymic medulla from C57BL/6 mice were sorted and their transcriptional profiles compared. Cells were gated as viable, Epcam+ CD45- for thyroid and viable, Epcam+ CD45- UEA+ for thymic medulla.
2018-05-01 | GSE71243 | GEO
Project description:We have compared the transcriptional profiles from three epithelial cell types to find common and differentially expressed genes. Epithelial cells from small intestine and kidney from C57BL/6 mice were sorted and their transcriptional profiles compared. Cells were gated as viable, Epcam+ CD45- for kidney, and viable, Epcam+ CD45- MHCIIhigh for small intestine MHCIIhi and viable, Epcam int CD45- MHCII low for small intestine MHCIIlo.
2018-05-01 | GSE68678 | GEO
Project description:To investigate gene expression profile of medullary thymic epithelial cells with high surface density of MHC class II (mTEChigh), a murine parent into F1 hematopoietic stem cell transplantation model was administered. As recipient mice (B6xDBA/2)F1 (B6D2F1) were used that received either a syngeneic transplant from a (B6xDBA/2)F1 mouse or an allogeneic transplant from a B6 mouse, which leads to acute graft-versus-host disease (aGVHD). mTEChigh (CD45-EpCAM+Ly51-UEA1+MHCIIhigh) are sorted by FACS.
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Project description:Separate populations of M cells have been detected in the follicle-associated epithelium of Peyerâs patches (PPs) and the villous epithelium of the small intestine, but the traits shared by or distinguishing the two populations have not been characterized. Our separate study has demonstrated that M cells are rare but M-like cells positive for lectin Ulex europaeus agglutinin-1 and our newly developed M cell-specific mAb NKM16-2-4 are induced by oral administration of cholera toxin in the duodenal villous epithelium. Here, we determined the gene expression of PP M cells, villous M-like cells, and intestinal epithelial cells (IECs) isolated by a novel approach using FACS. Specific expression of glycoprotein 2 (GP2) and MARCKS-like protein (MLP) by PP M cells and not villous M-like cells was confirmed by additional mRNA and protein analyses. Comprehensive gene profiling also suggested that villous M-like cells share traits of both PP M cells and IECs, a finding that is supported by their unique expression of specific chemokines. The genome-wide assessment of gene expression facilitates discovery of M cell-specific molecules and enhances the molecular understanding of M cell immunobiology. Experiment Overall Design: Epithelial cells in duodenal Peyer's patches or villi of BALB/c mice treated with or without cholera toxin were dissociated and stained with NKM16-2-4 monoclonal antibody (mAb)-FITC, lectin UEA-1-PE, and anti-CD45 mAb-APC-Cy7. Naive PP M cells (NKM16-2-4+, UEA-1+, and CD45- cells), CT-induced villous M-like cells (NKM16-2-4+, UEA-1+, and CD45- cells), and naive intestinal epithelial cells (NKM16-2-4-, UEA-1-, and CD45- cells) were isolated by FACS. Affymetrix GeneChip Mouse Genome 430 2.0 Array was used in triplicate for each cell fraction.
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Project description:Gene expression in different thymic stromal cells and subsets thereof was analyzed in 6-12 week old wild type (C57BL/6) and Aire knock-out (mixed background) mice. Thymic stromal cells were purified by sequential enzymatic digestion (collagenase, collagenase/dispase and trypsin) followed by gradient centrifugation and FACS sorting. Sort criteria were as follows: dendritic cells (CD11c+, F4/80 -), macrophages (F4/80+, CD11c-), cTECs (CD45–/lo, CDR1/Ly51+, Ep-CAM+) and mTECs (CD45–/lo, CDR1/Ly51–, Ep-CAM+). mTECs of wild-type and Aire knock-out mice were further subdivided according to CD80 expression levels. For microarray analysis total RNA from thymic stromal cell samples of two independent experiments was pre-amplified and biotinylated by two rounds of cDNA synthesis and in vitro transcription. Fluorescence readings were evaluated by using Microarray Suite 5.0 software. Keywords = thymus Keywords = promiscuous gene expression Keywords = AIRE Keywords: ordered
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Project description:The survival of isolated metastatic cells and expansion into macroscopic tumour has been recognized as a limiting step for metastasis formation in several cancer types yet the determinants of this process remain largely uncharacterized. In colorectal cancer (CRC), we identify a transcriptional programme in tumour-associated stromal cells, which is intimately linked to a high risk of developing recurrent disease after therapy. A large proportion of CRCs display mutational inactivation of the TGF-beta pathway but paradoxically they are characterized by high TGF-beta production. In these tumours, TGF-beta instructs a transcriptional programme in stromal cells, which confers a high risk of developing metastatic disease. We purified by FACS CD31(+), CD45(+), FAP(+) and Epcam(+) populations from fresh CRC samples and assessed their gene expression profiles Freshly obtained tumours from 6 CRC patients treated at Hospital del Mar or Hospital Clinic (Barcelona, Spain) were minced and incubated with Collagenase IV (100 U mL-1). Enzymatic reaction was stopped by adding 10% FBS, single cells were collected by sequential filtering and resuspended in ammonium chloride (0.15M) to lyse erythrocytes. Cells were stained with FAP unconjugated rabbit antibody. After two washes with HBSS, cells were stained with an APC conjugated anti-rabbit antibody; anti-hEPCAM/TROP1-FITC, anti-CD31-PE and anti-CD45-PE-Cy7 conjugated antibody. Dead cells were labelled with Propidium Iodide. Fluorescence Activated Cell Sorting (FACS) was used to separate 2000 cells from each population; [CD45(+), EPCAM(-), CD31(-), FAP(-)], [CD45(-) EPCAM(+), CD31(-), FAP(-)], [CD45(-), EPCAM(-), CD31(+),FAP(-)] and [CD45(-), EPCAM(-), CD31(-), FAP(+)].
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