|
| Status |
Public on Jan 01, 2011 |
| Title |
pateint 3 skin fibroblasts 0h post serum treatment |
| Sample type |
RNA |
| |
|
| Source name |
skin fibroblast, patient 3, serum + 0h
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: skin fibroblast
post serum time point: 0 hours
patient number: 3
|
| Treatment protocol |
Cells were seeded at a density of 1x104 cells/cm2 in 10cm diameter dishes, cultured in normal growth medium for 24 hours, then serum starved (0.1% FCS) for 48 hours prior to stimulation with 10% FCS. Samples were taken at 0h and 6h following serum stimulation of both oral mucosal and skin fibroblast cultures derived from each patient.
|
| Growth protocol |
Primary cell cultures (derived from 6mm biopsies) of oral mucosal (normal, non-diseased buccal mucosa) fibroblasts and patient-matched skin fibroblasts were grown under standard mammalian tissue culture conditions (37 degrees celcius and 5% carbon dioxide) in fibroblast-serum containing medium (F-SCM) as previously described (Stephens P, Davies KJ, al-Khateeb, et al. , J Dent Res, 1996; 75: 1358-64).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
TRIzol extraction of total RNA was performed according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
5 μg RNA was used to generate double-stranded cDNA by using a T7-linked oligo(dT) primer. Biotinylated cRNA was synthesised using the T7 RNA transcript labelling kit (Enzo Diagnostics, Farmingdale NY, USA)
|
| |
|
| Hybridization protocol |
10 μg fragmented cRNA was then hybridised overnight to the human HG-U133A GeneChip
|
| Scan protocol |
GeneChips were scanned using the Agilent Gene Array Scanner controlled by Microarray Suite 5.0.
|
| Description |
Gene expression data from human skin fibroblasts following serum starvation
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling (TGT 100) as normalization method.
|
| |
|
| Submission date |
May 04, 2010 |
| Last update date |
Jan 01, 2011 |
| Contact name |
Philip Stephens |
| E-mail(s) |
StephensP@cardiff.ac.uk
|
| Phone |
+44 (0)29 2074 2529
|
| URL |
http://www.cardiff.ac.uk/dentl/contactsandpeople/academicstaff/stephens-phil-dr-overview_new.html
|
| Organization name |
Cardiff University
|
| Department |
Dental School
|
| Lab |
Wound Biology Group
|
| Street address |
Dental Drive, Heath Park
|
| City |
Cardiff |
| State/province |
South Glamorgan |
| ZIP/Postal code |
CF14 4XY |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL96 |
| Series (1) |
| GSE21648 |
Gene expression in human oral mucosal fibroblasts and patient-matched skin fibroblasts |
|
