|
| Status |
Public on Feb 20, 2007 |
| Title |
hBSMC_vitamin D_rep1 |
| Sample type |
RNA |
| |
|
| Source name |
human bronchial smooth muscle cell_VitD_100nM_24h
|
| Organism |
Homo sapiens |
| Characteristics |
ethnicity: Hispanic
Gender: Male
Age: 31
Tissue: primary human bronchial smooth muscle cells
|
| Biomaterial provider |
Cambrex, East Rutherford, New Jersey
|
| Treatment protocol |
BSMCs were grown in 25 cm2 flasks in SmGM and starved for 24 hours (SmBM supplemented with 0.1% FBS) before stimulation. Supernatant was removed and cells were incubated for 24 hours in fresh growth factor-free medium with 100 nM of 1alpha,25-Dihydroxyvitamin D3 (Sigma, Saint Louis, Missouri).
|
| Growth protocol |
Cells were growth in Smooth muscle Growth Medium (SmGM-2 Bulletkit, Cambrex) which included Smooth muscle Basal Medium (SmBM) supplemented with 5 % Fetal Bovine Serum (FBS), hEGF (0.5 ng/ml), hFGF (2 ng/ml), insulin (5 µg/ml) and GA-1000 (100 ng/ml of Gentamicin and 0.1 ng/ml of Amphotericin-B), according to manufacturer's instructions.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were lysed by adding 2 ml of trizol reagent (Invitrogen) and homogenized by passing through a syringe with a needle (26G). RNA was picked up in the aqueous phase following phase separation by chloroform. The RNA was then precipitate with isopropanol, washed with ethanol (75%), dissolved in DEPC-treated RNAse/DNAse free water and quantified by spectrophotometry (Ultrospec 2100 pro).
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA was prepared according to the standard Affymetrix protocol (see complete protocol descriptions: http://www.affymetrix.com/support/technical/manual/expression_manual.affx)
|
| |
|
| Hybridization protocol |
Hybridization was performed according to the standard Affymetrix protocol (see complete protocol descriptions: http://www.affymetrix.com/support/technical/manual/expression_manual.affx)
|
| Scan protocol |
Arrays were scanned with the Affymetrix GeneChip® Scanner 3000 7G and the data was extracted using Microarray Analysis Suite 5.0 (Affymetrix).
|
| Description |
Human primary bronchial smooth muscle cells were grown to near confluence, starved for 24 hours (medium containing 0.1% FBS) and then treated with 100 nM of vitamin D (1alpha,25-Dihydroxyvitamin D3) for an additional 24 hours before RNA extraction.
|
| Data processing |
Expression values were extracted using the Robust Multichip Average method (Irizarry RA, Hobbs B, Collin F, et al. 2003. Exploration, normalization, and summaries of high density oligonucleotide array probe level data. Biostatistics 4: 249-64)
|
| |
|
| Submission date |
Jun 23, 2006 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Yohan Bossé |
| E-mail(s) |
[email protected]
|
| Organization name |
Laval University
|
| Street address |
2725, chemin Sainte-Foy
|
| City |
Quebec |
| State/province |
Quebec |
| ZIP/Postal code |
G1V 4G5 |
| Country |
Canada |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE5145 |
Genes regulated by vitamin D in bronchial smooth muscle cells |
|
| Relations |
| Reanalyzed by |
GSE49910 |
| Reanalyzed by |
GSE64985 |
| Reanalyzed by |
GSE119087 |
