|
| Status |
Public on Jun 04, 2012 |
| Title |
EndometrialStromalCell_Choco_Ectopic_2-H2O2 |
| Sample type |
RNA |
| |
|
| Source name |
ectopic endometrial stromal cell from patients with ovarian endometrioma, treated with H2O2
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: ectopic endometrial stromal
age: 32
cycle phase: treated with GnRH
diagnosis: endometriosis
r-afs stage(score): IV(45)
|
| Treatment protocol |
After 24 hours of incubation, medium was exchanged with Phenol Red-free DMEM(Sigma D5921) + 1uM hydrogen peroxide + 200IU/ml potassium benzylpenicillin(Meiji) + 200ug/ml streptomycin sulfate(Meiji) + 2.5mg/ml amphotericin B(Sigma A2942). After 60 hours of incubation, RNA was collected. Supplementary hydrogen peroxide was added to medium every 48 hours. Control cells were incubated in Phenol Red-free DMEM(Sigma D5921) + 200IU/ml potassium benzylpenicillin(Meiji) + 200ug/ml streptomycin sulfate(Meiji) + 2.5mg/ml amphotericin B(Sigma A2942) for 60 hours and its RNA was collected.
|
| Growth protocol |
Endometrial stromal cells were incubated in DMEM (Sigma D5921) + 10% FBS (BioWest) + 200IU/ml potassium benzylpenicillin (Meiji) + 200ug/ml streptomycin sulfate (Meiji) + 2.5mg/ml amphotericin B (Sigma A2942) for 24 hours.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using RNeasy Mini(QIAGEN).
|
| Label |
Cy3
|
| Label protocol |
According to the manufacturer’s protocol (One-Color Microarray-Based Gene Expression Analysis (Quick Amp Labeling), Version 5.7, March 2008).
|
| |
|
| Hybridization protocol |
According to the manufacturer’s protocol (One-Color Microarray-Based Gene Expression Analysis (Quick Amp Labeling), Version 5.7, March 2008).
|
| Scan protocol |
According to the manufacturer’s protocol (One-Color Microarray-Based Gene Expression Analysis (Quick Amp Labeling), Version 5.7, March 2008, Agilent Scanner Settings).
|
| Data processing |
Microarray data were analyzed following import into GeneSpring 11.5 software (Agilent Technologies)
Threshold: 1.0
Logbase: 2
Technology: Agilent.SingleColor.14850
Normalization: Shift to 75 percentile
Baseline Transformation: None
|
| |
|
| Submission date |
Aug 18, 2011 |
| Last update date |
Jun 04, 2012 |
| Contact name |
Shinya Toyokuni |
| E-mail(s) |
akatsuka@med.nagoya-u.ac.jp
|
| Organization name |
Nagoya University
|
| Department |
Pathology
|
| Street address |
65 Tsuruma-Cho, Showa-Ku
|
| City |
Nagoya |
| State/province |
Aichi |
| ZIP/Postal code |
466-8550 |
| Country |
Japan |
| |
|
| Platform ID |
GPL6480 |
| Series (1) |
| GSE31515 |
Transcriptome analysis of endometriotic stromal cells treated with hydrogen peroxide |
|
