|
| Status |
Public on Aug 28, 2010 |
| Title |
ASC_FBS_P2_rep5 |
| Sample type |
RNA |
| |
|
| Source name |
ASC, Passage 2, FBS
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: adipose tissue-derived mesenchymal stem cells
supplement: 10% fetal bovine serum (FBS)
|
| Treatment protocol |
Cells were cultured under three different culture-conditions: basal DMEM, low glucose, supplemented with 10 % fetal bovine serum, 10% human serum or 10% thrombin-activated platelet releasate plasma (for reference Kocaömer et al. Stem Cells. 2007;25(5):1270-8.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were harvested when reaching at least 80% confluency. At passage 2, extraction of RNA was performed for each sample using the RNeasy Mini Kit (Qiagen). The quality of RNA was confirmed by checking the RNA integrity number using using the RNA 6000 Nano LabChip Kit and Agilent 2100 Bioanalyzer (Agilent Technologies).
|
| Label |
cy3
|
| Label protocol |
lHSel protocol: RNA from each sample was amplified and labeled using a Low input RNA fluorescent linear amplification kit (Agilent).
|
| |
|
| Hybridization protocol |
An in situ hybridization kit plus (Agilent) was used as a hybridization buffer. After hybridization in a hybridization chamber for 16 h at 65°C, microarrays were washed in washing solution 1+2 (20 × SSC and 10% Triton X-102) for 10 min each and in Agilent stabilization and drying solution.
|
| Scan protocol |
Immediately after the washing process, microarrays were scanned by a GMS418 laser scanner (Affymetrix, Eurofins MWG GmbH) at an excitation wavelength of 543 nm (Cy3) and 633 nm (Cy5).
|
| Description |
Gene expression of ACS from Passage 2 cultured in FBS
|
| Data processing |
Fluorescent intensities of scanned images were quantified by ArrayVision Ver.7 (Imaging research) and normalized applying Loess normalization procedure to adjust the Cy3 and Cy5 channels. Differential gene expression analysis was performed using the One-way ANOVA with the commercial software package SAS JMP7 Genomics 3.1 (SAS Institute). A false positive rate of a=0.05 with FDR correction was taken as the level of significance.
|
| |
|
| Submission date |
Aug 27, 2010 |
| Last update date |
Aug 27, 2010 |
| Contact name |
Karen Bieback |
| E-mail(s) |
karen.bieback@medma.uni-heidelberg.de
|
| Organization name |
Institute of Transfusion Medicine and Immunology, Medical Faculty Mannheim, Heidelberg University
|
| Department |
Institute of Transfusion Medicine and Immunology
|
| Street address |
Ludolf-Krehl-Strasse 13-17
|
| City |
Mannheim |
| ZIP/Postal code |
68167 |
| Country |
Germany |
| |
|
| Platform ID |
GPL1708 |
| Series (1) |
| GSE23851 |
Altered Gene Expression in Human Adipose Stem Cells Cultured with Fetal Bovine Serum Compared to Human Supplements |
|
