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Bovine Glomerular and Aortic Endothelial Cells

ABSTRACT: SAGE libraries were prepared from mRNA of cultured bovine glomerular and aortic endothelial cells. Cells both cell lines were positive for acetylated LDL uptake and vWF expression, and were proven to be free of non-endothelial cells. Cultures were studied at passage 9-10. The cells were confluent, cultured in parallel, under identical conditions. Cells were plated on gelatin coated cell culture plastic in RPMI 1640 medium containing 10% FBS and streptomycin/penicillin. The last medium change was 48 hours prior to mRNA harvest. Keywords = endothelial, glomerular, kidney, aorta, Keywords: other

ORGANISM(S): Bos taurus

SUBMITTER: Barbara Ballermann

PROVIDER: E-GEOD-110 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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A SAGE-based comparison between glomerular and aortic endothelial cells.

Sengoelge Gürkan G Luo Wensheng W Fine Derek D Perschl Agnes M AM Fierlbeck Wolfgang W Haririan Abdolreza A Sorensson Jenny J Rehman Tausif-Ur TU Hauser Peter P Trevick Jacob S JS Kulak Stephen C SC Wegner Binytha B Ballermann Barbara J BJ

American journal of physiology. Renal physiology 20050118 6

Endothelial cells have many characteristics in common, but significant morphological and functional differences exist between endothelial cells from different anatomic sites. The specific glomerular endothelial (GEn) cell transcript repertoire is unknown. We sought to determine whether endothelial cells derived from bovine glomeruli display a distinct transcriptional profile compared with bovine aortic endothelium (BAE) under identical conditions. Serial analysis of gene expression (SAGE), which ...[more]

PMID: 15657302

Similar Datasets

Bovine Glomerular and Aortic Endothelial Cells

Project description:SAGE libraries were prepared from mRNA of cultured bovine glomerular and aortic endothelial cells. Cells both cell lines were positive for acetylated LDL uptake and vWF expression, and were proven to be free of non-endothelial cells. Cultures were studied at passage 9-10. The cells were confluent, cultured in parallel, under identical conditions. Cells were plated on gelatin coated cell culture plastic in RPMI 1640 medium containing 10% FBS and streptomycin/penicillin. The last medium change was 48 hours prior to mRNA harvest. Keywords = endothelial, glomerular, kidney, aorta, Keywords: other

2002-12-10 | GSE110 | GEO

SAGE analysis of human glomerular endothelial cell

Project description:Tissue from macroscopically healthy parts of nephrectomized kidneys. Endothelial cells were purified from the cell outgrowth of glomeruli twice using CD31 magnetic beads and cultured on fibronectin coated surface. Cells were not passaged more than eight times and stained with vWf, CD31, and after stimulation with TNF alpha also with CD62E/P. This GEO Series was created by the GEO staff as part of a cleanup effort to ensure that all GEO Samples are included within a Series entry.

2010-12-01 | E-GEOD-25760 | biostudies-arrayexpress

glomerular endotelium and EPC MVs

Project description:Glomerular endothelial cells were cultured in normal condition and treated or not with microvesicles derived from endothelial progenitor cells. mRNA profiling of glomerular endothelial cells , treated with MVs, was analyzed after normalization with mRNA profiling of untreated cells.

2012-11-27 | E-MEXP-3762 | biostudies-arrayexpress

SAGE analysis of human glomerular endothelial cell

2010-12-01 | GSE25760 | GEO

Primary human umbilical vein and arterial endothelial cells

Project description:Human umbilical cords were obtained from the Lucille Packard Children Hospital. In this study, we devised a rapid isolation scheme to preserve the in vivo phenotype of each endothelial subtype. ECs were first isolated from umbilical cords by collagenase perfusion through the vein or artery as described. Cells were further purified using a Percoll density gradient (Amersham Biosciences, Piscataway, NJ) to remove residual erythrocytes and platelets. 1-5 x 106 ECs were then cultured overnight on gelatin-coated T12.5 flasks in Clonetics EGM-2 media (Cambrex Bio Science, Walkersville, MD) in which the ECs generally reached confluence the next morning. The confluent EC were trypsinized and subjected to two rounds of immuno-magnetic beads purification, adapted from a published purification protocol. There was a negative selection step using CD14, CD45 and CD64 to remove residual contaminating leukocytes, followed by positive selection using a mouse anti-endothelial cell monoclonal antibody (anti-CD146/clone P1H12 purchased from Chemicon, Temecula, CA). Total processing time was limited to 20 to 24 hours. The homogeneous, viable, primary ECs were used immediately to construct the library. The construction of SAGE libraries was performed with the I-SAGE kit (Invitrogen, Carlsbad, CA) per manufacturer’s instructions Keywords: other

2009-12-30 | E-GEOD-1539 | biostudies-arrayexpress

Gene expression profiles of the diabetic glomerular endothelial cell

Project description:The objective of this study is to create an encyclopedia of all genes expressed in the glomerular endothelial cell under normal and diabetic conditions. We utilized Tie2-GFP transgenic mice to mark cells of the glomerular endothelium. To induce diabetic nephropathy (DB), a genetic model of DB, BKS.Cg-m +/+ Leprdb/J from Jax laboratories was used. We utilized fluorescent activated cell sorting (FACS) to isolate glomerular endothelial cells from normal and diabetic mice. The RNAs from these samples were isolated and utilized to hybridize to microarrays, which offers a powerful, efficient and effective method for the creation of a gene expression atlas. Microarrays were used to identify the transciptional differences that occur in the glomular endothelium of a diabetic mouse. Diabetic and control mice carrying the Tie2-GFP transgenic were utilized to isolate the endothelial cells from the adult glomerulus. The endothelial cells were isolated from the glomerulus using FACS. RNA was isolated and the gene expression profiles were determined by microarrays.

2010-08-16 | E-GEOD-21324 | biostudies-arrayexpress

Dynaimic changes in glomerular cells in the development of diabetes

Project description:To investigate dynamic changes in glomerular cells, including podocyte, mesangial cells and glomerular endothelial cells, in the development of diabetic nephropathy We then performed gene expression profiling analysis using data obtained from RNA-seq of glomerular cells of control(m/m), diabetic (db-/- 6-week-old) and diabetic nephropathy (db-/- 10-week-old with albuminuria) mice

2023-10-27 | GSE219268 | GEO

Gene expression profiles of the diabetic glomerular endothelial cell

2010-08-16 | GSE21324 | GEO

Tripartite separation of glomerular proteomes from reporter-free mice

Project description:Renal blood filtration occurs in a functional unit called the glomerulus. The resident cell types comprise the filtration barrier, namely podocytes, mesangial cells and glomerular endothelial cells. Here we introduce a glomerular cell isolation protocol, which enables the separation of these three cell types in sufficient amounts and purity to allow detailed protein-biochemical investigations. We demonstrate that the expression of fluorescent transgenes in glomerular cells can result in proteome artifacts. We show that different mouse strains have different glomerular cell type proteomes. Further, we demonstrate the power of the technique to identify new glomerular cell type-enriched proteins and demonstrate the potential of this globally applicable technique in the dissection of cell-specific disease responses and intra-glomerular cell-type crosstalk.

2021-11-12 | PXD024481 | Pride

Single-cell profiling of glomerular cells provides a dynamic picture in experimental diabetic kidney disease

Project description:Background: Recent single-cell RNA sequencing (scRNA-seq) analyses have offered much insight into cell-specific gene expression profiles in normal kidneys. However, in diseased kidneys, understanding of changes in specific cells, particularly glomerular cells, remains limited. Methods: To elucidate the glomerular cell–specific gene expression changes in diabetic kidney disease, we performed scRNA-seq analysis of isolated glomerular cells from streptozotocin-induced diabetic endothelial nitric oxide synthase (eNOS)–deficient (eNOS-/-) mice and control eNOS-/- mice. Results: We identified five distinct cell populations, including glomerular endothelial cells, mesangial cells, podocytes, immune cells, and tubular cells. Using scRNA-seq analysis, we confirmed the expression of glomerular cell–specific markers and also identified several new potential markers of glomerular cells. The number of immune cells was significantly higher in diabetic glomeruli compared with control glomeruli, and further cluster analysis showed that these immune cells were predominantly macrophages. Analysis of differential gene expression in endothelial and mesangial cells of diabetic and control mice showed dynamic changes in the pattern of expressed genes, many of which are known to be involved in diabetic kidney disease. Moreover, gene expression analysis showed variable responses of individual cells to diabetic injury. Conclusion: Our findings demonstrate the ability of scRNA-seq analysis in isolated glomerular cells from diabetic and control mice to reveal dynamic changes in gene expression in diabetic kidneys, with variable responses of individual cells. Such changes, which might not be apparent in bulk transcriptomic analysis of glomerular cells, may help identify important pathophysiologic factors contributing to the progression of diabetic kidney disease.

2019-03-08 | GSE127235 | GEO

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