|
| Status |
Public on Nov 11, 2013 |
| Title |
CD14-CD45-EpCAM+ cells In bone marrow, non liver metastasis group, replicate 7 |
| Sample type |
RNA |
| |
|
| Source name |
CD14-CD45-EpCAM+ cells In bone marrow, non liver metastasis group, replicate 7
|
| Organism |
Homo sapiens |
| Characteristics |
tissue: CD14-CD45-EpCAM+ cells
group: non liver metastasis
gender: male
age: 49y
|
| Treatment protocol |
Colorectal cancer patients were administered anesthesia and 20 mL BM was taken from the right and left anterior iliac crests before surgery. Mononucleated cells were collected using a standard Ficoll-Hypaque gradient technique. To enrich for EpCAM+ cells, CD14+ cells were removed from the whole bone marrow using auto MACSTM pro (Milteny Biotec, Bergisch Gladbach, Germany) with anti-CD14 immunomagnetic beads (clone; TÜK4, Milteny Biotec). Next, CD45+ cells were removed by treatment with anti-CD45 immunomagnetic beads (clone; 5B1; Milteny Biotec). The residual CD14−CD45− cells were then incubated with FcR blocking reagent (Milteny Biotec), followed by incubation with anti-EpCAM immunomagnetic beads (clone; HEA-125, Milteny Biotec), and the CD14−CD45−EpCAM+ cells were taken up.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated using the miRNeasy kit (Applied Biosystems, Carlsbad, CA) following the manufacturer’s protocol. Total RNA concentration and purity were assessed with a spectrophotometer. RNA was quantified using a NanoDrop-1000 spectrophotometer and quality was monitored with the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA).
|
| Label |
Cy3
|
| Label protocol |
100 ng totalRNA samples were labeled and hybridized with the miRNA Complete Labeling and Hyb Kit(Agilent, Santa Clara,CA,USA) in accordance with the manufacture's procedure.
|
| |
|
| Hybridization protocol |
Cy3-labeled samples were hybridized with the miRNA Complete Labeling and Hyb Kit(Agilent, Santa Clara,CA,USA) acording to the manufacture's procedure.
|
| Scan protocol |
Microarray slides were scanned with Agilent G2505C scanner. The sanned image files were analyzed using Agilent FeatureExtraction software ver 10.7.3.1.
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.7.3.1(Agilent) using default parameters (protocol miRNA_107_Sep09 and Grid:021827_D_F_20100604) to obtain background subtracted and spatially detrended Processed Signal intensities. The signal values were 90 percentile normalization with Agilent GeneSpring ver11.5.1
|
| |
|
| Submission date |
Oct 25, 2013 |
| Last update date |
Nov 11, 2013 |
| Contact name |
Hiroshi Takeyama |
| E-mail(s) |
[email protected]
|
| Phone |
81-6-6879-3251
|
| Organization name |
Graduate School of Medicine, Osaka University
|
| Department |
Gastroenterological Surgery
|
| Street address |
Yamada-oka 2-2
|
| City |
Suita |
| State/province |
Osaka |
| ZIP/Postal code |
565-0871 |
| Country |
Japan |
| |
|
| Platform ID |
GPL10850 |
| Series (1) |
| GSE51716 |
MicroRNA expression signatures of CD14-CD45-EpCAM+ cells in human bone marrow for colorectal cancer liver metastasis |
|
