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Expression data of mesenchymal cells from mouse liver

ABSTRACT: There are a few markers available to distinguish hepatic stellate cells (HSCs), portal fibroblasts (PFs), and mesothelial cells (MCs) in the adult mouse liver. To identify genes uniquely expressed in these cells, we isolated HSCs having Vitamin A lipids by FACS based on their autofluorescence. We isolated a mixed population containing PFs and MCs as Vitamin A lipid-negative and GFP-positive cells by FACS from Collagen1a1 promoter-driven GFP transgenic mice. MCs were isolated by FACS using anti-GPM6A antibodies from adult mouse livers. PFs were isolated by FACS as Vitamin A lipid-negative, GFP-positive, and GPM6A-negative cells. We also isolated HSCs, MCs, and PFs from Collagen1a1 promoter-driven GFP transgenic mouse livers having biliary fibrosis by FACS. Biliary fibrosis was induced by bile duct ligation for 3 weeks. Sham operation was performed as a control. After separation of these cells, we isolated total RNAs and analyzed mRNA expression by microarray analysis.

ORGANISM(S): Mus musculus

PROVIDER: GSE66788 | GEO | 2016/03/11

SECONDARY ACCESSION(S): PRJNA277966

REPOSITORIES: GEO

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Expression data of mesenchymal cells from mouse liver

Project description:There are a few markers available to distinguish hepatic stellate cells (HSCs), portal fibroblasts (PFs), and mesothelial cells (MCs) in the adult mouse liver. To identify genes uniquely expressed in these cells, we isolated HSCs having Vitamin A lipids by FACS based on their autofluorescence. We isolated a mixed population containing PFs and MCs as Vitamin A lipid-negative and GFP-positive cells by FACS from Collagen1a1 promoter-driven GFP transgenic mice. MCs were isolated by FACS using anti-GPM6A antibodies from adult mouse livers. PFs were isolated by FACS as Vitamin A lipid-negative, GFP-positive, and GPM6A-negative cells. We also isolated HSCs, MCs, and PFs from Collagen1a1 promoter-driven GFP transgenic mouse livers having biliary fibrosis by FACS. Biliary fibrosis was induced by bile duct ligation for 3 weeks. Sham operation was performed as a control. After separation of these cells, we isolated total RNAs and analyzed mRNA expression by microarray analysis. Total RNA was extracted with RNAqueous Micro (Ambion) and the probes for the microarray were synthesized using the Ovation RNA amplification system V2 and FL-Ovation cDNA Biotin module V2 (Nugen). The labeled probes were hybridized with GeneChip Mouse Genome 430 2.0 arrays (Affymetrix) and signals were analyzed with Genomic Suite software (Partek).

2016-03-11 | E-GEOD-66788 | biostudies-arrayexpress

Heterogeneity in the hepatic stellate cell population in mouse liver

Project description:We utilized two methods of hepatic stellate cell (HSC) isolation, which leverage distinct properties of these cells, including their vitamin A content and collagen expression. HSCs were isolated either from wild type (WT) mice in the C57BL/6 genetic background by flotation in a Nycodenz density gradient, followed by fluorescence activated cell sorting (FACS) based on vitamin A autofluorescence, or from collagen-green fluorescent protein (GFP) mice by FACS based on GFP expression, from a GFP transgene driven by the collagen I promoter. Array analysis was conducted on the Affymetrix Mouse Genome 430 2.0 Array (Affymetrix, Santa Clara, CA).

2011-09-01 | E-MEXP-3231 | biostudies-arrayexpress

Expression data from developing mouse livers

2013-02-04 | GSE39064 | GEO

RNAseq comparison of gene expression profiles in ScxGFP positive cells and ScxGFP negative cells from E13.5 forelimb samples

Project description:RNAs were isolated from FACS sorted ScxGFP positive cells and GFP negative cells of forelimbs at E13.5, and characterized by RNAseq

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Transcriptional profiling of hepatocyte, reprogrammed hepatocyte, and biliary cells

Project description:Following liver injury, a subset of hepatocytes adopt features of biliary epithelial cells (BECs) in a process known as biliary reprogramming. The aim of this study is to elucidate the molecular events accompanying this dramatic shift in cellular identity. Wild type hepatocytes, wild type biliary cells, DDC-treated hepatocytes, DDC-treated biliary cells, and DDC-treated reprogrammed cells were isolated by FACS, RNA isolated, and sequenced.

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Expression data from developing mouse livers

Project description:Roles of mesothelial cells (MCs) are poorly understood during liver development and injury. We identified podoplanin (Pdpn) as a cell surface markers for mesothelial cells in E12.5 mouse developing liver. To identify genes uniquely expressed in MCs, we isolated MCs from E12.5 mouse livers by FACS using anti-Pdpn antibodies and performed microarray analysis. The E12.5 liver was digested with trypsin-EDTA and incubated with antibodies against Pdpn. MCs were isolated as Pdpn+ population by FACS. Total RNA was extracted with RNAqueous Micro (Ambion) and the probes for the microarray were synthesized using the Ovation RNA amplification system V2 and FL-Ovation cDNA Biotin module V2 (Nugen). The labeled probes were hybridized with GeneChip Mouse Genome 430 2.0 arrays (Affymetrix) and signals were analyzed with Genomic Suite software (Partek).

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Bulk RNA-seq of Sox9-deficient intrahepatic biliary epithelial cells

Project description:To define the regulatory role of Sox9 in intrahepatic biliary epithelium, we conducted bulk RNA-seq on biliary epithelial cells (BECs) from adult control (Sox9fl/fl) and conditional Sox9 knockout (Sox9cKO; Sox9fl/fl:Albumin-Cre) mice. Livers were dissociated to single cells and BECs isolated by FACS based on CD326 positivity. CD31+ and CD45+ cells were excluded by FACS. Gallbladder and extrahepatic tissues were excluded by dissection. RNA was isolated using Ambion RNAqueous Micro kit and libraries were made using Takara SMARTer Stranded Total RNA-seq Kit v3 Pico.

2024-01-01 | GSE249385 | GEO

Transcription factor induction of vascular blood stem cell niches in vivo [ATAC-Seq 2]

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2023-03-23 | GSE227113 | GEO

Targets of EOMES in mouse CD4+ T cells

Project description:To gain mechanistic insights into how EOMES regulates CD4+ T-cell differentiation and function, we performed an RNA-seq analysis using Eomes-GFP reporter mice (Eomes+/GFP) to isolate GFP+ (EOMES+) and GFP- (EOMES-) CD4+ T-cells by FACS sorting. We also sequenced RNA from Eomes-deficient GFP+ and GFP- CD4+ T-cells isolated from EomesΔT/GFP knock-out mice, in which one Eomes allele is disrupted by GFP insertion and the DNA-binding domain of the other allele is deleted in T cells by Lck-driven Cre recombinase. Hence, we had two populations of GFP+ cells where the EOMES locus was transcribed, one with a transcriptionally active EOMES protein (Eomes+/GFP) and another one with no transcriptionally active EOMES (EomesΔT/GFP), plus the respective GFP-negative controls isolated from the same mice. A homogeneous population of EOMES-expressing CD4+ T cells were obtained by transferring naïve sorted CD25- CD45RBhigh CD4+ T-cells isolated from Eomes+/GFP reporter or EomesΔT/GFP knock-out donor animals into Rag2-/- mice. After three weeks of adoptive transfer GFP+ and GFP- CD4+ T-cell populations from these mice were FACS-sorted for transcriptome analysis by RNA sequencing.

2020-10-01 | GSE145145 | GEO

MiRNA expression in embryonic murine hepatocytes versus murine cholangiocytes at E18.5

Project description:Differentially expressed miRNAs between hepatocytes (GFP-neg in SOX9-GFP mice) and cholangiocytes (GFP-pos in SOX9-GFP mice) in mice at E18.5. We want to compare miRNA population in biliary cells and parenchymal cells (depleted from hematopoietic cells) during liver development. Cholangiocytes specifically express the transcription factor Sox9. This enables to separate cholangiocytes from the rest of the liver parenchyma by FACS from Sox9-GFP transgenic mice. In conclusion, we want to compare miRNA population in our GFP+ cells (cholangiocytes) and our GFP- cells (liver parenchyma depleted from hematopoietic cells).

2018-12-20 | GSE100145 | GEO

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