|
| Status |
Public on Jan 01, 2016 |
| Title |
A_1 |
| Sample type |
RNA |
| |
|
| Source name |
Cell culture, replicate 1
|
| Organism |
Mus musculus |
| Characteristics |
strain: C57BL/6
age: 8-12 week old mice
tissue: heart left atrial appendage
|
| Growth protocol |
Adult (8-12 weeks old) C57BL/6 mice were deeply anesthetized with isoflurane and left atrial appendages were removed. The explants were then digested three times (5min) with trypsin & collagenase D. Strong enzyme digestion was performed with trypsin 0.25% and collagenase D 0.1% and alternatively, weak enzyme digestion with trypsin 0.05% and collagenase D 0.1%. After digestion the explants were placed on 24 –well plates (tissue culture treated) in culture medium (CEM: Isocove’s modified Dulbecco’s Medium with 25mM HEPES, supplemented with 10% FBS (fetal bovine serum), 1% L-glutamine, 1% penicillin-streptomycin & 0.1mM 2 -mercaptoethanol).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA of cultured cells was extracted with High Pure RNA isolation kit (Roche) according to the manufacturer’s instructions. RNA -sample quality and integrity was tested with Bioanalyzer 2100 (Agilent Technologies). RNA from control tissue samples was isolated using Tri-reagent.
|
| Label |
Cy3
|
| Label protocol |
200 ng of total RNA for each sample was amplified and labeled with a fluorescent dye (Cy3 ) using the Low RNA Input Linear Amplification & Labeling kit (Agilent ).
|
| |
|
| Hybridization protocol |
The Cy3 labeled cRNA was hybridized to the 8 X 60K Agilent Whole Mouse Genome Oligo Microarray V2 (design 028005 Agilent Technologies, USA) for 17 hours at 65°C in an Agilent DNA-Microarray Hybridization Oven.
|
| Scan protocol |
The Microarray was scanned using the Agilent High-Resolution Microarray Scanner.
|
| Description |
Primary cells (c-kit+/CD45- sorted) from left atrial appendage tissue culture
A
A samples were treated with strong enzyme digestion.
|
| Data processing |
Data was extracted from the resulting images using the Feature Extraction 10.7 software.
|
| |
|
| Submission date |
Apr 24, 2014 |
| Last update date |
Jan 01, 2016 |
| Contact name |
Jussi Leinonen |
| E-mail(s) |
jussi.leinonen@mac.com
|
| Organization name |
Hadassah-Hebrew University Medical Center
|
| Department |
Cardiovascular Research Center, Heart Institute
|
| Street address |
Kiryat Hadassah p.o.b 12000
|
| City |
Jerusalem |
| ZIP/Postal code |
91120 |
| Country |
Israel |
| |
|
| Platform ID |
GPL13912 |
| Series (2) |
| GSE57064 |
Macrophage Precursor Cells from the Left Atrial Appendage of the Adult Heart Spontaneously Reprogram into a C-kit+/CD45- Stem Cell-Like Phenotype [Array analysis] |
| GSE57126 |
Macrophage Precursor Cells from the Left Atrial Appendage of the Adult Heart Spontaneously Reprogram into a C-kit+/CD45- Stem Cell-Like Phenotype |
|
