|
| Status |
Public on Jul 08, 2006 |
| Title |
Bronchial epithelial cells, day 8, donor 3 |
| Sample type |
RNA |
| |
|
| Source name |
Primary human bronchial epithelial cells were obtained from three different healthy, nonsmoking adult donors by cytological brushing at bronchoscopy
|
| Organism |
Homo sapiens |
| Characteristics |
Cells were harvested at day 8 of air-liquid interface culture
|
| Growth protocol |
Cells were obtained by cytological brushing at bronchoscopy and expanded to passage three in bronchial epithelial growth medium (BEGM, Clonetics). Cells were plated on vitrogen-coated filter supports inserted into 12-well culture plates. 500 nM retinoic acid was added to culture medium after cells reached 100% confluence to promote differentiation. Air-liquid interface (ALI) culture was initiated 48 hours later by removing the apical medium, and basolateral media containing 100nM retinoic acid was used for the ALI culture period.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were lysed and total RNA was prepared for microarray analysis using the RNeasy kit (Qiagen), as per the manufacturerÆs protocol.
|
| Label |
biotin
|
| Label protocol |
Biotin-labeled cRNA was synthesized using the ENZO BioArrayÖ HighYieldÖ RNA Transcript Labeling Kit (T7)
|
| |
|
| Hybridization protocol |
Fragmented cRNA was hybridized to the microarray according to the Affymetrix GeneChip« Expression Analysis Technical Manual (November 2004).
|
| Description |
Bronchial epithelial cells, day 8, donor 3 , Primary human bronchial epithelial cells were obtained from three different healthy, nonsmoking adult donors by cytological brushing at bronchoscopy
|
| Data processing |
Affymetrix .cel files were processed at Expression Analysis (Durham, NC) using the REDI (REduction of Invariant Probes) Analysis to remove data from poorly performing probes in combination with Robust Multichip Average (RMA) analysis, which performs background correction, probe-level normalization and probe-set summary.
|
| |
|
| Submission date |
Jul 07, 2006 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Andrea Ross |
| E-mail(s) |
ross.andrea@epa.gov
|
| Organization name |
US EPA
|
| Street address |
NHEERL/HSD/CRB
|
| City |
Research Triangle Park |
| State/province |
NC |
| ZIP/Postal code |
27711 |
| Country |
USA |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE5264 |
Human bronchial epithelial cell differentiation time course |
|
| Relations |
| Reanalyzed by |
GSE64985 |
| Reanalyzed by |
GSE119087 |
