|
| Status |
Public on Jul 11, 2007 |
| Title |
Murine embyonic pancreas stem cells |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
Total RNA from murine CD133negPDGFRb- cells labeled with Cyanine-5 (red).
|
| Organism |
Mus musculus |
| Characteristics |
Pancreas
|
| Treatment protocol |
To isolate CD133highPDGFRb- cells and CD133negPDGFRb- cells, 500 of embryonic day 13.5 fetal pancreases were obtained, digested in 0.04 units Liberase Blenzyme 3 (Roche) at 37℃ for 5 min, then triturated in low glucose DMEM media containing 0.2% bovine serum albumin. After staining with monoclonal antibodies, these cells were subjected to FACS cell sorting and each fraction was collected to extract total RNA.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNAs were extracted using RNAeasy Micro Kit (QIAGEN) following manufacturer’s instructions.
|
| Label |
Cy5
|
| Label protocol |
Sample labeling was done using Agilent’s Low RNA Fluorescent Linear Amplification Kit PLUS (5188-5340: Agilent) following manufacture’s instructions. Amplified cRNAs were quantified using the Agilent 2100 bioanalyzer and RNA 6000 Nano kit.
|
| |
|
| Channel 2 |
| Source name |
Total RNA from murine CD133highPDGFRb- cells labeled with Cyanine-3 (green).
|
| Organism |
Mus musculus |
| Characteristics |
Pancreas
|
| Treatment protocol |
To isolate CD133highPDGFRb- cells and CD133negPDGFRb- cells, 500 of embryonic day 13.5 fetal pancreases were obtained, digested in 0.04 units Liberase Blenzyme 3 (Roche) at 37℃ for 5 min, then triturated in low glucose DMEM media containing 0.2% bovine serum albumin. After staining with monoclonal antibodies, these cells were subjected to FACS cell sorting and each fraction was collected to extract total RNA.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNAs were extracted using RNAeasy Micro Kit (QIAGEN) following manufacturer’s instructions.
|
| Label |
Cy3
|
| Label protocol |
Sample labeling was done using Agilent’s Low RNA Fluorescent Linear Amplification Kit PLUS (5188-5340: Agilent) following manufacture’s instructions. Amplified cRNAs were quantified using the Agilent 2100 bioanalyzer and RNA 6000 Nano kit.
|
| |
|
| |
| Hybridization protocol |
Performed using Gene Expression Hybridization Kit (5188-5242 : Agilent)
|
| Scan protocol |
Scanned on an Agilent G2565BA scanner.
Images were quantified using Agilent Feature Extraction Software (version 9.1.3).
|
| Description |
CD133negPDGFRb- cells _CD133highPDGFRb- cells
|
| Data processing |
Dye biases between two signals, Red and Green, were corrected using Lowess normalization, subtracted background and converted to log10 of processed Red signal/processed Green signal.
|
| |
|
| Submission date |
May 15, 2007 |
| Last update date |
May 15, 2007 |
| Contact name |
Yuichi Hori |
| E-mail(s) |
horiy@med.kobe-u.ac.jp
|
| Phone |
+81-78-382-5371
|
| Fax |
+81-78-382-5379
|
| Organization name |
Kobe University School of Medicine
|
| Department |
Surgery
|
| Street address |
7-5-1 Kusunoki-cho, Chuo-ku
|
| City |
Kobe |
| State/province |
Hyogo |
| ZIP/Postal code |
650-0017 |
| Country |
Japan |
| |
|
| Platform ID |
GPL4134 |
| Series (1) |
| GSE7800 |
Identification of genes enriched in putative stem/progenitor cells from mouse embryonic pancreas |
|
