|
| Status |
Public on Aug 23, 2010 |
| Title |
Clone_354_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
thymic epithelial cells 4months after second transplatation into skin, bulk 354
|
| Organism |
Rattus norvegicus |
| Characteristics |
strain: Sprague-Dawley rats which constitutively express the Enhanced Green Fluorescent protein (EGPF) (green rat CZ-004” SDTgN (act-EGFP)obsCZ-004)
|
| Treatment protocol |
TECs anf hair follicle clones were cultivated on a feeder layer of irradiated 3T3-J2 cells in a 3:1 DMEM F12 medium supplemented as described (Oshima et al., Cell 2001).
|
| Growth protocol |
Thymic lobes were removed from embryos at E16, and from postnatal rats at P1, P7, and P137. Dissected thymic lobes were harvested and kept in cold Hank’s Balanced Salt Solution (HBSS) complemented with 10% Fetal Bovine Serum during dissection. Single lobes were placed in individual 35 mm size Petri dishes and washed in HBSS without Ca2+ and Mg2+ twice before being dissociated in 0.05% trypsin solution containing 0.1% EDTA at 37°C. Skin grafts containing GFP-positive hair follicles were carefully dissected from the back of recipient nude mice and dissociated in 30 ml of 0.05% trypsin containing 0.1% EDTA at 37°C for 60 to 120 min.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was prepared using the RNeasy Mini Kit (QIAGEN Inc.)
|
| Label |
biotin
|
| Label protocol |
100 nanogram of total RNA were amplified and labeled using Affymetrix whole transcript sense target labeling protocol (kits #900673, #900671)
|
| |
|
| Hybridization protocol |
Affymetrix Rat Gene 1.0 ST arrays were hybridized with 5 microgramme of labeled, amplified cDNA , washed, stained and scanned according to the protocol described in Affymetrix GeneChip® Expression Analysis Manual (Fluidics protocol FS450_0007)
|
| Scan protocol |
Scanning was done on an Affymetrix GeneChip Scanner 7G
|
| Data processing |
Normalized expression signals were calculated from Affymetrix CEL files using RMA (Irizarry et al., 2003) in Affymetrix Expression Console software.
|
| |
|
| Submission date |
May 05, 2010 |
| Last update date |
Aug 23, 2010 |
| Contact name |
Sylvain Pradervand |
| E-mail(s) |
Sylvain.Pradervand@unil.ch
|
| Phone |
+41 21 692 39 08
|
| Organization name |
UNI Lausanne
|
| Department |
CIG
|
| Lab |
DNA Array Facility
|
| Street address |
Genopode
|
| City |
Lausanne |
| ZIP/Postal code |
1015 |
| Country |
Switzerland |
| |
|
| Platform ID |
GPL6247 |
| Series (1) |
| GSE21686 |
Comparison of thymic epithelial cells and hair follicle stem cells |
|
