|
| Status |
Public on May 16, 2012 |
| Title |
Senescent HUVEC 3 |
| Sample type |
RNA |
| |
|
| Source name |
Umbilical Vein Endothelial Cells
|
| Organism |
Homo sapiens |
| Characteristics |
age: senescent (XIII passage)
cell type: HUVEC
|
| Treatment protocol |
Endothelial cell replicative senescence was studied by subjecting endothelial cells to subsequent passages until XIII
|
| Growth protocol |
HUVEC cells were purchased from Clonetics Corporation (Lonza, Basel, Switzerland) and cultured in endothelial growth medium EGM-2 (Lonza)
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with RNA purification kit (Norgen Biotek Corporation, Thorold, ON, Canada)
|
| Label |
FAM
|
| Label protocol |
About 150-200 ng of total RNA were converted to cDNA by priming with a mixture of looped primers using manufacturer’s instructions (MegaPlex kit, Applied Biosystems, Foster City, CA). Pre-amplification was performed using 3 microliters of input RNA with PreAmp kit (Applied Biosystem, Foster City, CA). 9 microliters of pre-amplified cDNA were used for mature miRs profiling by real-time PCR instrument equipped with a 348 well reaction plate (7900 HT, Applied Biosystem, Foster City CA) and human MiR Array pool A
|
| |
|
| Hybridization protocol |
n/a
|
| Scan protocol |
n/a
|
| Description |
HUVEC XIII passage
|
| Data processing |
The relative amount of each miRNA was calculated using the comparative threshold (Ct) method with ΔCt = Ct(miRNA)-Ct(RNU44). We used median normalization method for array data. Relative quantification of miRNA expression was calculated with the 2-ΔΔCt method. ΔΔCt was calculated as CT microRNA- median Ct values obtained in the sample profiling.
|
| |
|
| Submission date |
May 14, 2012 |
| Last update date |
May 16, 2012 |
| Contact name |
Fabiola Olivieri |
| E-mail(s) |
f.olivieri@univpm.it
|
| Phone |
+390712206242
|
| Fax |
+390712206240
|
| Organization name |
Università Politecnica delle Marche
|
| Department |
Dept of clinical and molecular sciences
|
| Lab |
Experimental Pathology
|
| Street address |
Via Tronto
|
| City |
Ancona |
| State/province |
Ancona |
| ZIP/Postal code |
60020 |
| Country |
Italy |
| |
|
| Platform ID |
GPL14851 |
| Series (1) |
| GSE37972 |
Real-time quantitative PCR analysis of Human Umbilical Vein Endothelial Cells (HUVEC) |
|
