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Steroids Effect on HUVEC Response to LPS or Cytokines

ABSTRACT: Pooled first passage cryopreserved, human umbilical vein endothelial cells (HUVEC) were obtained from Cascade Biologics (Portland, OR), thawed, and cultured according to the supplier’s recommendations in Medium 200. After thawing, cells were seeded onto 0.2% gelatin coated tissue culture plates and cultivated under 95% air, 5% CO2 at 37 oC. Confluent cells were cultured in steroids-depleted medium for 96 hrs before the addition of 17β-estradiol (17βE) or hydrocortisone (H) to 1 µM for 5 hrs. For Lipopolysaccharide (LPS) or cytokine mix (CM)-treatments, cells or pretreated with vehicle (0.1% ethanol), 1µM 17βE, or H for 1 hr were exposed to 100 ng/ml LPS or a mixture of proinflammatory cytokines (CM, 500 U/ml IL-1b, 2500 U/ml TNF-α, and 1250 U/ml IFN-γ) for 4 hours. All treatments were done in quadruplicates. At the end of treatment, total cellular RNA was isolated. Keywords: parallel sample

ORGANISM(S): Homo sapiens

PROVIDER: GSE1486 | GEO | 2004/06/19

SECONDARY ACCESSION(S): PRJNA90761

REPOSITORIES: GEO

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Steroids_Effect_on_HUVEC_Response_to_LPS_or_Cytokines

Project description:Pooled first passage cryopreserved, human umbilical vein endothelial cells (HUVEC) were obtained from Cascade Biologics (Portland, OR), thawed, and cultured according to the supplierâs recommendations in Medium 200. After thawing, cells were seeded onto 0.2% gelatin coated tissue culture plates and cultivated under 95% air, 5% CO2 at 37 oC. Confluent cells were cultured in steroids-depleted medium for 96 hrs before the addition of 17Î²-estradiol (17Î²E) or hydrocortisone (H) to 1 ÂµM for 5 hrs. For Lipopolysaccharide (LPS) or cytokine mix (CM)-treatments, cells or pretreated with vehicle (0.1% ethanol), 1ÂµM 17Î²E, or H for 1 hr were exposed to 100 ng/ml LPS or a mixture of proinflammatory cytokines (CM, 500 U/ml IL-1b, 2500 U/ml TNF-Î±, and 1250 U/ml IFN-Î³) for 4 hours. All treatments were done in quadruplicates. At the end of treatment, total cellular RNA was isolated.

2010-06-09 | E-GEOD-1486 | biostudies-arrayexpress

Gene expression of murine B cells activated via TLR4 or TLR4 + CD40: Group1 naM-CM-/ve B cells, Group2 LPS activated B cells 24h, Group3 LPS + anti-CD40 activated B cells 24h, Group4 LPS activated B cells 72h, and Group5 LPS + anti-CD40 activated B cells 72h.

Project description:Murine B cells can be activated via the surface receptors TLR4 and CD40. For a global assessment of differences in gene expression between these two different modes of B cell activation a genome wide transcriptome analysis was performed. In order to dissect different gene expression profiles of B cells, activation was induced by LPS or LPS + anti-CD40 for 24h and 72h. Both activation states were compared to each other but also to naM-CM-/ve B cells. Gene expression profiles of naM-CM-/ve B cells, B cells activated with LPS and B cells activated with LPS + anti-CD40. Affymetrix MG 430 2.0 whole genome arrays were performed in quadruplicates for naM-CM-/ve B cells, B cells activated via TLR4 for 24 h and 72 h, and B cells activated via TLR4 + CD40 for 24 h and 72 h (20 arrays in total). To obtain genes significantly regulated upon stimulation with LPS, the expression profiles of naM-CM-/ve B cells, B cells activated with LPS for 24 h, and B cells activated with LPS for 72 h were compared to each other. After total RNA extraction, reverse transcription, cDNA extraction, the biotinylated cRNA was transcribed, fragmented, and 15 M-BM-5g cRNA hybridized in quadruplicates for each of the five groups to the 20 GeneChip arrays: Group1 naM-CM-/ve B cells, Group2 LPS activated B cells 24h, Group3 LPS + anti-CD40 activated B cells 24h, Group4 LPS activated B cells 72h, and Group5 LPS + anti-CD40 activated B cells 72h. Access to PDF: http://dx.doi.org/10.1038/nature12979 All chips were imported into BioRetis (http://www.bioretis-analysis.de) and are open to the community now. If you want to obtain one or more lists of HPCDA significant genes you should click on a link of single chips (e.g. GSM879034) to get a description how to manage this.

2014-02-24 | E-GEOD-35998 | biostudies-arrayexpress

Therapeutic intervention of monocytes to LPS challenge

Project description:To investigate the possible therapeutic effects of ibuprofen and Pep19-2.5 alone or in combination on the LPS-response of human monocytes. Human monocytes were challenged with 0.1ng/ml LPS and/or 10ng/ml Pep19-2.5 LPS. Medium controls served as unstimulated samples. LPS and Pep19-2.5 was used either alone or together to LPS-challenged samples. RNA was extracted after 4 hrs of stimulation. Three biological replicates were conducted

2015-07-02 | E-GEOD-65855 | biostudies-arrayexpress

LPS time-course in WT and Stk40 KO BMDMs

Project description:Bone marrow was harvested from Rosa26CreER; Stk40+/+ (WT; n = 3) and Rosa26CreER; Stk40loxp/loxp (Stk40 KO; n = 3) mice and differentiated for 6 days in the presence of 100 nM 4-OHT to generate WT and Stk40 KO bone-marrow derived macrophages (BMDMs). 2. On day 7 following differentiation BMDMs were treated with 100 ng x ml-1 LPS and harvested at 0 hrs, 6 hrs, 16 hrs, and 32 hrs following LPS exposure. 3. The cells were snap-frozen at the time of harvest. RNA was extracted using the Qiagen RNeasy mini kit as per manufacturer’s protocol including the on-column DNase digestion. Groups: There are cells from 3 mice x 2 genotypes x 4 time points G1: WT 0 hr LPS G2: WT 6 hr LPS G3: WT 16 hr LPS G4: WT 32 hr LPS G5: Stk40 KO 0 hr LPS G6: Stk40 KO 6 hr LPS G7: Stk40 KO 16 hr LPS G8: Stk40 KO 32 hr LPS

2018-12-28 | GSE124410 | GEO

MicroRNA profiling by array of mouse skeletal muscle cells treated with TNF-alpha or IGF-I

Project description:Murine skeletal muscle cells pmi28 were cultured in growth medium (HamM-^Rs F10, supplemented with 20 % FCS, 2 mM L-glutamine, and 1 % Penicillin / Streptomycin). Myoblasts were cultured on laminin-1 coated dishes for an additional 24 h before switching a fraction of dishes to differentiation medium (DMEM medium containing 2 % horse serum, 2 mM L glutamine, and 1 % Penicillin / Streptomycin) and to differentiation medium containing either 2 x 103 U/ml mouse recombinant TNF-? (Roche Applied Science), or 5 M-5g/ml mouse recombinant IGF-I (Sigma-Aldrich), or carrier only. pmi28 cells (TNF-? and IGF-I treatments as well as controls) were harvested 24 h after the induction of differentiation. The experiments were performed in quadruplicates or triplicates, respectively.

2013-05-15 | E-MTAB-1114 | biostudies-arrayexpress

Phagosome proteome of activated murine macrophage

Project description:Proteomics of carboxylated polystyrene bead (1.0 um) phagosomes from murine bone marrow-derived macrophages. cells were either resting or treated with 100 U/ml IFN-γ (PeproTech) and 100 ng/ml LPS (Sigma) for 24 h, 20 ng/ml Interleukin-4 (IL4) (BD Pharmingen) for 48 h, 20 ng/ml Interleukin-13 (IL13), 10 ng/ml Interleukin-10 (IL10)for 48 h or Reprogrammed (IL4 was incubated with BMDMs for 24 h, and the medium was replaced with fresh medium containing IFN-γ/LPS to incubate for another 24 h). Phagosomes were isolated after 30 min bead inoculation.

2019-03-22 | PXD004520 | Pride

Transcriptome sequencing wide functional analysis of human mesenchymal stem cells (LPS)

Project description:Using RNA-seq, we report here that BM-MSC cells have a distinct transcriptomic signature and express a unique cluster of transcripts in response to 4 hrs LPS (1ug/ml).

2019-04-03 | GSE97723 | GEO

A549 parenchymal cell activation

Project description:Alveolar epithalial cells (A549) were used as a model of pulmonary parenchymal cell activation to quiery gene expression profile of activation with either cyclic stretch (20% elongation), TNF (20 ng/ml), LPS (1 mcg/ml) or TNF+STRETCH at both 1 and 4 hrs. Keywords: time-course

2004-10-03 | GSE1541 | GEO

The activation of inflammatory response genes by LPS is blocked by HDAC inhibitor Trichostatin A (TSA)

Project description:TSA treatment blocks LPS-induction of several inflammatory target genes in primary macrophages. The microarray experiment was performed to identify LPS-inducible, LXR-sensitive target genes that retained LPS-induction in the presence of TSA. Thioglycollate-elicited macrophages were isolated from mice by peritoneal lavage 3 days following peritoneal injection of 2.5 ml 3% thioglycollate (DIFCO). Cells were plated in RPMI medium 1640 and 10% fetal bovine serum, washed after 5 h the medium was removed and cells were fed with fresh medium containing 0.5% fetal bovine serum. Cells were treated with TSA (Wako) at 100nM and/or with LPS (Sigma) at concentration of 100 ng/ml in the absence or presence of receptor-specific agonist for LXR (GW3965) at 1 µM. Keywords: TSA, LPS, LXR agonist

2006-12-15 | GSE6352 | GEO

B-cell activating factor (BAFF) stimulation of Burkitt Lymphoma cell line [RNA-Seq]

Project description:RNA-Seq profiling of Burkitt Lymphoma cell line (BL2) with B-cell activating factor (BAFF) for 24 hrs . The Burkitt Lymphoma cell line were either only cultured in cell culture medium supplemented with 10 mM HEPES at 1 × 106 cells/ml or additionally incubated with B-cell activating factor (BAFF) for 24 hrs

2018-05-23 | GSE99768 | GEO

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