|
| Status |
Public on Nov 19, 2013 |
| Title |
SGBS_12h_1 |
| Sample type |
RNA |
| |
|
| Source name |
SGBS cell line undergoing differentiation
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: SGBS
time: 12 h
|
| Treatment protocol |
See growth protocol for induction of differentiation.
|
| Growth protocol |
SGBS cells were cultured in Dulbecco's modified Eagle's medium (DMEM)/Nutrient Mix F12 (Gibco) containing 8 mg/l biotin, 4 mg/l pantothenate, 0.1 mg/mg streptomycin and 100 U/ml penicillin (OF medium) supplemented with 10% FBS in a humidified 95%air/5%CO2 incubator. The cells were seeded into culture medium flasks or plates, which were coated with a solution of 10 microL/ml fibronectin and 0.05% gelatine in phosphate-buffered saline. Confluent cells were cultured in serum-free OF medium for 2 days followed by stimulation to differentiate with OF media supplemented with 0.01 mg/ml human transferrin, 200 nM T3, 100 nM cortisol, 20 nM insulin, 500 microM IBMX and 100 nM rosiglitazone (Cayman Chemicals). After day 4, the differentiating cells were kept in OF media supplemented with 0.01 mg/ml human transferrin, 100 nM cortisol and 20 nM insulin.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using TriSure (Bioline). 1 mL of TriSure was added per a confluent 6-well to lyse the cells. RNA extracted with 200 uL chloroform and precipitated from the aqueous phase with 400 uL isopropanol by incubating at -20C overnight.
|
| Label |
Cy3
|
| Label protocol |
Labelled as recommended by manufacturer
|
| |
|
| Hybridization protocol |
Hybridized as recommended by manufacturer
|
| Scan protocol |
Scanned as recommended by manufacturer
|
| Description |
SGBS preadipocyte cells originate from male patient with SGB syndrome and are cultured 2 days in serum-free OF medium prior to differentiation. Subsequently QuickDiff medium is added for the first 4 days of differentiation and then changed for 3FC medium. Please see Wabitsch M. et al. Int J Obes Relat Metab Disord. 2001 for more details
|
| Data processing |
R/Bioconductor lumi package with vst transformation and rsn normalization. (control probe values in separate table, subtracted prior to vst transformation using lumiB method bgAdjust)
|
| |
|
| Submission date |
Oct 04, 2012 |
| Last update date |
Nov 19, 2013 |
| Contact name |
Merja Heinäniemi |
| Organization name |
university of eastern finland
|
| Street address |
Yliopistonranta 1
|
| City |
Kuopio |
| ZIP/Postal code |
70210 |
| Country |
Finland |
| |
|
| Platform ID |
GPL6947 |
| Series (2) |
| GSE41352 |
Integrated analysis of transcript level regulation of metabolism during human adipocyte differentiation [time course] |
| GSE41578 |
Integrated analysis of transcript level regulation of metabolism during human adipocyte differentiation |
|
