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| Status |
Public on Mar 07, 2018 |
| Title |
The effects of testosterone and lipopolysaccharide on gene expression in immortalized ocular surface and adnexal cells |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Immortalized human corneal, conjunctival, and meibomian gland epithelial cells were pre-treated with dihydrotesteosterone (DHT) or vehicle, then exposed to lipopolysaccharide (LPS) and ligand binding protein (LBP) or vehicle, lysed, and subjected to microarray analysis of RNA expression.
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| Overall design |
Cells were cultured in keratinocyte serum-free medium (KSFM) supplemented with bovine pituitary extract (BPE; 25 ug/mL HCEC and HCjE, 50 ug/mL HMGEC), 5 ng/mL epidermal growth factor (EGF), penicillin and streptomycin. On reaching confluence, cells were rinsed twice with a phosphate-buffered saline solution and exposed to a stratification medium consisting of DMEM and F12 (Mediatech, Inc) with 10% fetal bovine serum, epidermal growth factor (10 ng/mL), penicillin, and streptomycin for 2 days. After this time period, cells were incubated in serum-free DMEM/F12 and exposed to vehicle (1% bovine serum albumin [BSA]), or LPS (15 μg/ml) and LBP (150 ng/ml), for six hours. The LPS and LBP were dissolved in DMEM and the BSA in PBS.
For androgen-related studies, the stratification medium contained 10% charcoal and dextran-treated fetal bovine serum with ethanol or DHT (10nM); the poststratification medium also contained ethanol or DHT.
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| Contributor(s) |
Sullivan DA, Sahin A, Darabad RR |
| Citation(s) |
29763693 |
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| Submission date |
Mar 06, 2018 |
| Last update date |
Jan 06, 2020 |
| Contact name |
David A. Sullivan |
| E-mail(s) |
david.sullivan@schepens.harvard.edu
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| Phone |
617-912-0287
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| Organization name |
Schepens Eye Research Institute
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| Lab |
David A. Sullivan
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| Street address |
20 Staniford St.
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| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02114 |
| Country |
USA |
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| Platforms (1) |
| GPL10558 |
Illumina HumanHT-12 V4.0 expression beadchip |
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| Samples (36)
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| Relations |
| BioProject |
PRJNA437166 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE111496_RAW.tar |
106.1 Mb |
(http)(custom) |
TAR (of IDAT) |
| Processed data included within Sample table |
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