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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.

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