|
| Status |
Public on Dec 31, 2006 |
| Title |
Simvastatin treated 24h, sample 2, Simvastatin is anti-inflammatory in macrophages |
| Sample type |
RNA |
| |
|
| Source name |
human peripheral blood monocyte-derived macrophages
|
| Organism |
Homo sapiens |
| Characteristics |
from unknown healthy blood-donor volunteers (Finnish Red Cross)
|
| Biomaterial provider |
Finnish Red Cross
|
| Treatment protocol |
24h before sample collection cells were stimulated with 10 um simvastatin in serum free medium.
|
| Growth protocol |
HPBM Cells were isolated from buffy coats using Ficoll-Paque gradient centrifugation (as described Pietarinen-Runtti, et al. Am.J Physiol Cell Physiol. 2000;278:C118-125). During isolation, monocytes from 3 individuals were pooled. Adherent cells were cultured in standard medium (20% human serum, Cambrex for differentiation into macrophages. The macrophage-phenotype at day 7 after isolation was confirmed with the typical shape of macrophages and also by macrophage-immunostaining (mAB CD68, DAKO, Denmark, dilution 1:200), where macrophages presented >95% of the cell population. 36 hours prior to sample collection cell growth medium was changed to serum free medium.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from the cells using Trizol reagent (Gibco BRL, USA) according to manufacturers’ instructions. The quality and amount of RNA was assessed by spectrophotometry (NanoDrop, USA) and by agarose gel electrophoresis.
|
| Label |
Biotin
|
| Label protocol |
cDNAs were prepared from RNAs (5 ug of RNA) with reverse transcriptase (Superscript II primed by a poly (T) oligomer/T7 promoter). cDNAs were subsequently used as a template to make biotin-labeled cRNA with an in vitro transcription reaction according to Affymetrix standard protocol.
|
| |
|
| Hybridization protocol |
cRNAs (15 ug) were hybridized to Affymetrix HGU133 Plus 2.0 oligonucleotide arrays, which was processed and scanned according to manufacturer’s instructions.
|
| Scan protocol |
Standard Affymetrix protocoll.
|
| Description |
-
|
| Data processing |
Affymetrix GeneChip® Operating Software (GCOS) analysis
|
| |
|
| Submission date |
May 22, 2006 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Tiina Tuomisto |
| E-mail(s) |
tiina.tuomisto@uku.fi
|
| Organization name |
A.I.Virtanen Institute
|
| Department |
Molecular Medicine
|
| Lab |
SYH
|
| Street address |
Neulaniementie 2
|
| City |
Kuopio |
| ZIP/Postal code |
70210 |
| Country |
Finland |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE4883 |
Simvastatin has an anti-inflammatory effect on macrophages via upregulation of Kruppel-like factor-2 |
|
| Relations |
| Reanalyzed by |
GSE64985 |
| Reanalyzed by |
GSE119087 |
