|
| Status |
Public on Jun 28, 2012 |
| Title |
WJ-MSC 12vs4 exp5 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
WJ-MSC 12th cycle
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: WJ-MSC obtained from human umbilical cord at 12th cycle of expansion in vitro
cell pool: UCB
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from two WJCs pools at 12th in vitro cycle expansion using a total of 1000000 cells using the SVtotal RNA Izolation System kit (Promega, Madison, WI, USA)
|
| Label |
Cy3
|
| Label protocol |
Extracted RNA was linearly amplified using the Amino Allyl MessageAmp™ II aRNA Amplification Kit (Ambion, Austin, TX, USA). Five to ten ug of amplified aRNA were fluorescently labelled with Cy3
|
| |
|
| Channel 2 |
| Source name |
WJ-MSC 4th cycle
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: WJ-MSC obtained from human umbilical cord at 4th cycle of expansion in vitro
cell pool: UCB
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from two WJCs pools at 4th in vitro cycle expansion using a total of 1000000 cells using the SVtotal RNA Izolation System kit (Promega, Madison, WI, USA)
|
| Label |
Cy5
|
| Label protocol |
Extracted RNA was linearly amplified using the Amino Allyl MessageAmp™ II aRNA Amplification Kit (Ambion, Austin, TX, USA). Five to ten ug of amplified aRNA were fluorescently labelled with Cy5
|
| |
|
| |
| Hybridization protocol |
2 µg of aRNA fragmented and respectively Cy3-Cy5 labeled from each sample was hybridized on Phalanx Human OneArray™ by 1.5X OneArray hybridization Buffer at 42°C in oven for 14-16 hrs
|
| Scan protocol |
After hybridization, Cy3-Cy5 fluorescent signals were captured by a Confocal Laser Scanner "ScanArray Express" (Packard BioScience) and analysed using the software "ScanArray Express - MicroArray Analysis System" version 3.0 (Perkin Elmer)
|
| Data processing |
The values of the median signal intensity from each spot were subtracted from the local median background intensity. For each slide after local background subtraction, a LOWESS algorithm was used for row data normalization to evaluate signal to noise ratio and generate log ratios of sample vs. reference signal. A gene was considered to be differentially expressed when showing an absolute value of log-ratio higher or equal to 0.5, an index that translates to a fold-change of 1.4 in transcript quantity
|
| |
|
| Submission date |
Jan 09, 2012 |
| Last update date |
Jun 28, 2012 |
| Contact name |
Valentina Gatta |
| E-mail(s) |
v.gatta@unich.it
|
| Phone |
00390871541479
|
| Fax |
00390871541479
|
| Organization name |
G. d’Annunzio University Foundation Chieti
|
| Department |
Aging Research Center (CESI)
|
| Lab |
Department of Medical Genetics
|
| Street address |
Via Colle dell'Ara
|
| City |
Chieti |
| State/province |
Abruzzo |
| ZIP/Postal code |
66100 |
| Country |
Italy |
| |
|
| Platform ID |
GPL6254 |
| Series (1) |
| GSE34929 |
Gene expression profile of Wharton-jelly mesenchymal stem cells at 12th in vitro expansion cycle compared to Wharton-jelly mesenchymal stem cells at 4th in vitro expansion cycle |
|
