|
| Status |
Public on Oct 23, 2015 |
| Title |
PCBM1655, Passage 3, Technical Replicate 2 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
MSC Bone Marrow Derived
|
| Organism |
Homo sapiens |
| Characteristics |
donor id: PCBM1655
gender: FEMALE
cell type: human multipotent stromal cells (MSCs) derived from bone marrow
|
| Treatment protocol |
MSCs were passaged after reaching 80% confluency as determined by one sole cell biologist
|
| Growth protocol |
MSCs were grown in alpha minimum essential media with 10% FBS, 1% L-gluatmine & 1% penicillin G and streptomycin under humidified atmosphere of 5% CO2 at 37C
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted by the Qiagen Rneasy Mini kit
|
| Label |
Hyper5
|
| Label protocol |
Individual RNA material was labeled with Hyper5 and reference material (P3star or P3*) was labeled with Cy3. 5 micrograms RNA used as starting material. See manuscript for more detailed description
|
| |
|
| Channel 2 |
| Source name |
Reference Material (P3star): created from equal portions of total RNA from the 6 donors at passage 3
|
| Organism |
Homo sapiens |
| Characteristics |
sample type: Pooled reference Sample
rna source: equal portions of total RNA from 6 donors (PCBM1641 P3, PCBM1632 P3, PCBM1662 P3, 8F3560 P3, 167696 P3, 110877 P3).
|
| Treatment protocol |
MSCs were passaged after reaching 80% confluency as determined by one sole cell biologist
|
| Growth protocol |
MSCs were grown in alpha minimum essential media with 10% FBS, 1% L-gluatmine & 1% penicillin G and streptomycin under humidified atmosphere of 5% CO2 at 37C
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted by the Qiagen Rneasy Mini kit
|
| Label |
Cy3
|
| Label protocol |
Individual RNA material was labeled with Hyper5 and reference material (P3star or P3*) was labeled with Cy3. 5 micrograms RNA used as starting material. See manuscript for more detailed description
|
| |
|
| |
| Hybridization protocol |
Hyper5 and Cy3 labeled samples were mixed and hybridized on he Maui hybridization system for 16 hours at 42C
|
| Scan protocol |
Microarray slides were scanned using the 4000B GenePix Axon Scanner. PMT voltages of both channels were adjusted such that their intensity histograms overlap.
|
| Data processing |
LOESS/Linear normalization was performed using ArrayTrack software (FDA) with no background subtraction
|
| |
|
| Submission date |
Oct 23, 2015 |
| Last update date |
Oct 23, 2015 |
| Contact name |
Raj K Puri |
| E-mail(s) |
raj.puri@fda.hhs.gov
|
| Phone |
240-402-9171
|
| Organization name |
FDA
|
| Street address |
10903 New Hampshire Ave
|
| City |
Silver Spring |
| State/province |
Maryland |
| ZIP/Postal code |
20993 |
| Country |
USA |
| |
|
| Platform ID |
GPL18503 |
| Series (2) |
| GSE56362 |
Human Multipotent Stromal Cells: Passage 3 vs. Passage 5 vs. Passage 7 |
| GSE74303 |
Identification of Predictive Gene Markers for Multipotent Stromal Cell Proliferation |
|
