|
| Status |
Public on Oct 31, 2012 |
| Title |
Disc Cells in 3D_TNF-a (154) |
| Sample type |
RNA |
| |
|
| Source name |
Disc Tissue
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: Disc Tissue
tissue grade: IV
cell type: annulus cells
agent: TNF-a
|
| Treatment protocol |
They were fed 3 times per week for 9 days with minimal essential medium with 20% fetal bovine serum (MEM20) +/- 103 pM TNF-a. Cells were allowed to grow an additional 5 days without changing the media.
|
| Growth protocol |
Human annulus disc cells (100,000) were seeded into each piece (~0.5 cm3) of a collagen construct (Gelfoam; Pharmacia & Upjohn Co., Kalamazoo, MI). They were fed 3 times per week for 9 days with minimal essential medium with 20% fetal bovine serum (MEM20) +/- 10e3 PM TNF-a. Cells were allowed to grow an additional 5 days without changing the media.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Each piece of Gelfoam was homongenized and RNA isolated using TRIzol reagent
|
| Label |
biotin
|
| Label protocol |
Biotin-labeled cRNA were prepard according to the Affymetrix protocol for IVT labeling.
|
| |
|
| Hybridization protocol |
Fragmented cRNA was hybridized to the X3P chip in the Affymetrix hybridiztion buffer for 16 hrs at 45oC. X3P chips were washed and labeled in the Affymetrix Fluidics Station.
|
| Scan protocol |
GeneChips were scanned once via the Affymetrix 3000G scanner.
|
| Data processing |
The data were analyzed with the GCOS Affymetrix GeneChip Operating System (version 1.2) and Microarray Suite version 5.0 (MAS 5.0). Affymetrix default analysis settings and global scaling as normalization were used. Further normalization was acheived using GC-RMA (robust multiarray average) via GeneSifter analysis software (Viz Labs LLC, Seattle , WA; http://www.genesifter.net).
|
| |
|
| Submission date |
Oct 28, 2012 |
| Last update date |
Oct 31, 2012 |
| Contact name |
Helen Gruber |
| E-mail(s) |
[email protected]
|
| Organization name |
Carolinas HealthCare System
|
| Department |
Orthopaedic Surgery
|
| Lab |
Orthopaedic Research
|
| Street address |
1542 Graden Terrace
|
| City |
Charlotte |
| State/province |
NC |
| ZIP/Postal code |
28203 |
| Country |
USA |
| |
|
| Platform ID |
GPL1352 |
| Series (1) |
| GSE41883 |
Human annulus disc cells grown in a 3D collagen sponge and exposed to TNF-a |
|
