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Expression data from mammary epithelial cells harvested from 8-9 week old C3H female mice - various isolation conditions

ABSTRACT: Numerous lineage analysis studies have used FACS sorting as a separation technique prior to measuring mRNA expression patterns. To assess if FACS sorting causes non-specific changes in gene expression, we collected a heterogeneous population of mammary epithelial cells from three biological replicates. We then performed gene expression analysis on each replicate either prior to trypsin digestion, immediately following trypsin digestion, or following both the trypsin digestion and a mock FACS sort. In this dataset, we include gene expression data obtained from isolated murine mammary epithelial cells either prior to trypsin digestion, immediately following trypsin digestion, or following the trypsin digestion and a mock FACS sort.

ORGANISM(S): Mus musculus

PROVIDER: GSE61167 | GEO | 2014/09/05

SECONDARY ACCESSION(S): PRJNA260385

REPOSITORIES: GEO

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Expression data from mammary epithelial cells harvested from 8-9 week old C3H female mice - various isolation conditions

Project description:Numerous lineage analysis studies have used FACS sorting as a separation technique prior to measuring mRNA expression patterns. To assess if FACS sorting causes non-specific changes in gene expression, we collected a heterogeneous population of mammary epithelial cells from three biological replicates. We then performed gene expression analysis on each replicate either prior to trypsin digestion, immediately following trypsin digestion, or following both the trypsin digestion and a mock FACS sort. In this dataset, we include gene expression data obtained from isolated murine mammary epithelial cells either prior to trypsin digestion, immediately following trypsin digestion, or following the trypsin digestion and a mock FACS sort. Nine total samples were analyzed (3 biological replicates of three experimental conditions). Using LIMMA packages gene-wise comparisons were made between untrypsinized and trypsinized replicates as well as between trypsinized and mock FACS sorted replicates using and Pâ¤0.05 and a â¥1.5-fold difference between conditions.

2014-09-05 | E-GEOD-61167 | biostudies-arrayexpress

Single cell analysis of circulating B lymphocytes and myocardial B lymphocytes in naïve C57/B6 mice

Project description:Perfused naïve murine hearts harbor a sizeable population of B lymphocytes. In order to assess any differences between "myocardial" B cells and circulating B cells we FACS sorted B cells collected from the peripheral blood via cheek bleed and B cells isoalted from the heart after enzymatic digestion. CD45+, CD19+, Aqua- cells were facs sorted into PBS - with 0.04% BSA and processed for single cell RNA sequencing. Peripheral blood was colelcted immediately prior to sacrificing mice in a CO2 chamber. the blood and heart of 3 10 weeks old female mice was pooled together prior to FACS sorting.

2019-12-24 | GSE142515 | GEO

Transcription profiling of mouse thymic stromal cells from 6-12 week old wild type (C57BL/6) and Aire knock-outs (mixed background)

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 (CD45â/lo, CDR1/Ly51+, Ep-CAM+) and mTECs (CD45â/lo, CDR1/Ly51â, 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.

2007-12-15 | E-GEOD-2585 | biostudies-arrayexpress

promiscuous gene expression in the mouse thymus

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

2005-04-28 | GSE2585 | GEO

Proteomic profiling of FAC-sorted ex vivo and in vitro expanded Sca-1+cells derived from murine skeletal muscle

Project description:Stem cells antigen-1 (Sca-1) is an 18-kDa mouse glycosyl phosphatidylinositol-anchored cell surface protein that has widely been used for isolation of murine hematopoietic stem cells (HSCs). However, recently several studies have reported the expression of this protein in non-HSCs such as the liver, skin and muscles among others. In the current study, we used fluorescence-activated cell sorting (FACS) technique to sort Sca-1+ stem cells from mouse hindlimb muscles. The Sca-1+ cells were divided into two sets first set of Sca-1+ cells were processed for protein extraction immediately after sorting (ex vivo set) while the second set of samples, referred to as in vitro set, were expanded in cell culture after sorting and harvested after two passage and then processed for protein extraction. A high-resolution proteome map of the ex vivo and in vitro sets was generated Orbitrap Fusion Tribrid Mass Spectrometer. In total, the analysis led to the identification of 5,581 protein groups.

2021-06-21 | PXD022247 | Pride

Promiscuous gene expression in human medullary thymic epithelial cell subsets

Project description:Different human mTEC subsets (MUC1, CEACAM5 and SGLT1) were purified by sequential enzymatic digestion (collagenase/dispase, trypsin) followed by enrichment using magnetic beads (CD45 beads, Miltenyi Biotech) and FACS sorting. Cells of the surface phenotype CD45-, CDR2-, EpCAM+ were further subdivided into MUC1+/MUC1-, CEACAM5+/CEACAM5- and SGLT1+/SGLT1- fractions. RNA was isolated using μMACS™ SuperAmp™ protocol (Miltenyi Biotec) and hybridized to Illumina Whole-Genome Expression Beadchips. Gene expression of Antigen-positive and Antigen-negative mTEC subsets was compared.

2013-08-31 | GSE49625 | GEO

Transcriptional profiling of indolent and aggressive mammary lesions

Project description:We performed transcriptional profiling of tumor cells from indolent and aggressive lesions isolated from a mouse model of breast cancer. To initiate formation of mammary lesions, we delivered lentiviral particles carrying constitutively-activated Erbb2 tagged with HA and GFP (caErbB2) through the lactiferous glands of adult virgin female. Mammary glands bearing lesions were visualized under a fluorescence stereoscope, and regions harboring aggressive (>2mm, comprised of invasive lesions) and indolent (<2mm, enriched for in situ lesions) lesions were dissected away from each other. Lesions from multiple animals were pooled to obtain sufficient material for downstream analysis. Following tissue digestion, GFP-expressing epithelial cells were isolated by fluorescence-activated cell sorting (FACS), and then subjected to bulk RNA-seq.

2021-06-17 | GSE162005 | GEO

Single-cell transcriptional profiling of indolent and aggressive mammary lesions

Project description:We performed transcriptional profiling of tumor cells from indolent and aggressive lesions isolated from a mouse model of breast cancer. To initiate formation of mammary lesions, we delivered lentiviral particles carrying constitutively-activated Erbb2 tagged with HA and GFP (caErbB2) through the lactiferous glands of adult virgin female. Mammary glands bearing lesions were visualized under a fluorescence stereoscope, and regions harboring aggressive (>2mm, comprised of invasive lesions) and indolent (<2mm, enriched for in situ lesions) lesions were dissected away from each other. Lesions from multiple animals were pooled to obtain sufficient material for downstream analysis. Following tissue digestion, GFP-expressing epithelial cells and CD45+ immune cells were isolated by fluorescence-activated cell sorting (FACS), and then subjected to single-cell RNA-sequencing.

2021-06-17 | GSE161983 | GEO

Promiscuous gene expression in human medullary thymic epithelial cell subsets

2013-08-31 | E-GEOD-49625 | biostudies-arrayexpress

Gene expression profiles of the adult mesangial cell (GUDMAP Series_id: 37)

Project description:The long term objective is to create an encyclopedia of the expression levels of all genes in multiple components of the developing kidney. The central thesis is straightforward. The combination of fluorescent activated cell sorting (FACS) plus microarray analysis offers a powerful, efficient and effective method for the creation of a global gene expression atlas of the developing kidney. Microarrays with essentially complete genome coverage can be used to quantitate expression levels of every gene in FACS isolated components of the developing kidney. The ensuing rapid read-out provides an expression atlas that is more sensitive, more economical and more complete than would be possible by in situ hybridizations alone. Meis1-GFP BAC transgenic mice were utilized to isolate the mesangial cells from adult kidneys. The mesangial cells were isolated from the glomerulus using collagenase digestion, differential sieving, trypsin treatment and FACS. The RNA was isolated from purified mesangial cells and the gene expression profiles were determined by microarrays.

2010-03-12 | E-GEOD-20687 | biostudies-arrayexpress

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