|
| Status |
Public on Aug 13, 2012 |
| Title |
In vitro differentiated adipocytes_subject 91_day 8 |
| Sample type |
RNA |
| |
|
| Source name |
Isolated preadipocytes from subcutaneous adipose tissue, middle differentiation
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: subcutaneous adipose tissue
cell type: preadipocytes during in vitro differentiation to adipocytes
gender: female
age: 39
time of differentiation: day 8
subject: 91
|
| Treatment protocol |
Isolation, culture and differentiation of cells were performed exactly as described in PMID: 15492368.
|
| Growth protocol |
Subcutaneous adipose tissue was obtained from healthy subjects undergoing cosmetic liposuction (n=12). There was no selection for age, sex or BMI. From the tissue, we isolated and in vitro differentiated preadipocytes to adipocytes.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the cells at day 4/5 (early), 8 (middle) and 12 (late) of differentiation using the Nucleospin RNA11 kit (Macherey-Nagel, Duren, Germany). RNA concentration and purity were measured spectrophotometrically using a Nanodrop ND-1000 Spectrophotometer (Thermo Fisher Scientific), and high quality RNA was confirmed using the Agilent 2100 Bioanalyzer (Agilent Technologies, Palo Alto, CA).
|
| Label |
Biotin
|
| Label protocol |
Total RNA were amplified and biotin labeled using the Whole Transcript (WT) Sense Target Labeling Protocol.
|
| |
|
| Hybridization protocol |
From non-degraded high-quality total RNA, we prepared and hybridized biotinylated complementary RNA to Gene 1.0 ST Arrays, and then washed, stained and scanned slides using standardized protocols (Affymetrix Inc., Santa Clara, CA).
|
| Scan protocol |
We used standardized Affymetrix protocols (Affymetrix Inc., Santa Clara, CA).
|
| Description |
91_8
|
| Data processing |
Pre-processing was performed using the Affymetrix Expression Console version 1.1 and the following settings: Summarization: PLIER; Background Correction: PM-GCBG; Normalization: Global Median.
|
| |
|
| Submission date |
Dec 07, 2010 |
| Last update date |
Aug 14, 2012 |
| Contact name |
Ingrid Dahlman |
| E-mail(s) |
[email protected]
|
| Organization name |
Karolinska
|
| Street address |
Karolinska Huddinge
|
| City |
Stockholm |
| ZIP/Postal code |
14186 |
| Country |
Sweden |
| |
|
| Platform ID |
GPL6244 |
| Series (2) |
| GSE25402 |
Adipose Tissue MicroRNAs as Regulators of CCL2 Production in Human Obesity |
| GSE25910 |
Adipose Tissue MicroRNAs as Regulators of CCL2 Production in Human Obesity (differentiation data) |
|
