|
| Status |
Public on Feb 09, 2012 |
| Title |
pt #96 (Healthy Normal), Medium (unstimulated) |
| Sample type |
RNA |
| |
|
| Source name |
Human ASM, healthy, 2hr unstimulated control condition
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: bronchial airway smooth muscle (ASM) cells
treatment: none
time point: 2 hrs
patient identifier: 96
gender: male
age: 40
smoker status: ex-smoker
health status: healthy
fev1: 5.03
fev1%pred: 131
pc20: >16
therapy: none
atopy: non-atopic
allergen: none
|
| Treatment protocol |
Confluent human bronchial ASM cells were growth-arrested by FBS-deprivation for 48h in DMEM media containing 5µg/ml of human recombinant insulin, 5µg/ml of human transferrin, 5ng/ml of selenium (ITS) and antibiotics. Cells were then stimulated in fresh FBS-free media with or without recombinant human IL-17 [10ng/ml] for 2 hours.
|
| Growth protocol |
Human bronchial ASM cells were obtained by deep endobronchial biopsy from the right middle or lower lobe bronchi. ASM cells were grown at 37°C with 5% CO2 in Dulbecco's Modified Eagle Medium (DMEM) supplemented with 10% FBS, 100U/ml penicillin and 100µg/ml streptomycin. For all experiments, human ASM cells were used between passages 3 and 5.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted by the Qiagen RNeasy method, and RNA purity and integrity were evaluated by electrophoretic trace with the Agilent 2100 Bioanalyzer.
|
| Label |
biotin
|
| Label protocol |
Technical and platform specific procedures for the genome-wide, DNA-based whole transcript (WT) sense target (ST) Affymetrix HuGene v1.0 GeneChip were contracted to the University of Health Network Microarray Centre (UHNMC) Affymetrix Service.
|
| |
|
| Hybridization protocol |
Technical and platform specific procedures for the genome-wide, DNA-based whole transcript (WT) sense target (ST) Affymetrix HuGene v1.0 GeneChip were contracted to the University of Health Network Microarray Centre (UHNMC) Affymetrix Service.
|
| Scan protocol |
Technical and platform specific procedures for the genome-wide, DNA-based whole transcript (WT) sense target (ST) Affymetrix HuGene v1.0 GeneChip were contracted to the University of Health Network Microarray Centre (UHNMC) Affymetrix Service.
|
| Description |
96M
|
| Data processing |
The quality control metrics were evaluated by the Affymetrix Expression Console software v1.1.1 and the expression data were normalized using the Robust Multichip Analysis (RMA) algorithm. Secondary analysis such as statistical filtering of expression results and the creation of gene lists was performed with the Partek Genomics Suite v6.3.
|
| |
|
| Submission date |
Feb 08, 2012 |
| Last update date |
Feb 10, 2012 |
| Contact name |
Abdelilah S Gounni |
| E-mail(s) |
[email protected]
|
| Phone |
(204) 975-7750
|
| URL |
http://umanitoba.ca/faculties/medicine/units/immunology/soussigounni.html
|
| Organization name |
University of Manitoba
|
| Department |
Department of Immunology
|
| Street address |
750 McDermot Avenue
|
| City |
Winnipeg |
| State/province |
Manitoba |
| ZIP/Postal code |
R3E 0T5 |
| Country |
Canada |
| |
|
| Platform ID |
GPL6244 |
| Series (1) |
| GSE35643 |
Expression data from human bronchial airway smooth muscle (ASM) cells |
|
