|
| Status |
Public on May 01, 2011 |
| Title |
mesoangioblasts treated with 1 µM S1P at 24h |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
mesoangioblasts treated with 1 µM S1P at 24h
|
| Organism |
Homo sapiens |
| Characteristics |
developmental stage: adult
tissue: muscle biopsy
cell type: mesoangioblasts
|
| Treatment protocol |
For smooth muscle differentiation experiments, cells were seeded in p35 plates and when 90% confluent were shifted to DMEM without serum containing 1 mg/ml BSA and incubated in the presence or absence of 5 ng/ml TGFβ1 or 1 µM S1P (2 mM stock solution in dimethylsulfoxide). In some experiments cells were treated with 10 μM SKI-2, 60 min before agonist challenge.
|
| Growth protocol |
Human mesoangioblasts, isolated from adult muscle biopsies, were routinely grown in Mega Cell Sigma supplemented with 5% heat-inactivated FCS, 5 ng/ml bFGF, 2 mM L-glutamine, 100 U/ml penicillin and 100 μg/ml streptomycin, 1 μg/ml ciprofloxacin (Applichem, GmbH, Darmstadt) at 37°C in 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted following manufacturer's instructions.
|
| Label |
Cy5
|
| Label protocol |
Labeling reaction was performed following the protocols provided by Agilent according to the Two color microarray-based gene expression analysis manufacturer's protocol (Agilent).
|
| |
|
| Channel 2 |
| Source name |
mesoangioblasts Control at 24h
|
| Organism |
Homo sapiens |
| Characteristics |
developmental stage: adult
tissue: muscle biopsy
tissue: mesoangioblasts
|
| Treatment protocol |
For smooth muscle differentiation experiments, cells were seeded in p35 plates and when 90% confluent were shifted to DMEM without serum containing 1 mg/ml BSA and incubated in the presence or absence of 5 ng/ml TGFβ1 or 1 µM S1P (2 mM stock solution in dimethylsulfoxide). In some experiments cells were treated with 10 μM SKI-2, 60 min before agonist challenge.
|
| Growth protocol |
Human mesoangioblasts, isolated from adult muscle biopsies, were routinely grown in Mega Cell Sigma supplemented with 5% heat-inactivated FCS, 5 ng/ml bFGF, 2 mM L-glutamine, 100 U/ml penicillin and 100 μg/ml streptomycin, 1 μg/ml ciprofloxacin (Applichem, GmbH, Darmstadt) at 37°C in 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted following manufacturer's instructions.
|
| Label |
Cy3
|
| Label protocol |
Labeling reaction was performed following the protocols provided by Agilent according to the Two color microarray-based gene expression analysis manufacturer's protocol (Agilent).
|
| |
|
| |
| Hybridization protocol |
Hybridization and washing of the array were performed following the protocols provided by Agilent according to the Two color microarray-based gene expression analysis manufacturer's protocol (Agilent).
|
| Scan protocol |
Scanned on an Agilent G2565AA scanner
|
| Description |
Biological replicate 2 of 4
|
| Data processing |
Agilent Feature Extraction Software (v 8.5.1.1) was used for background subtraction and LOWESS normalization.
|
| |
|
| Submission date |
Aug 11, 2009 |
| Last update date |
May 01, 2011 |
| Contact name |
Alberto Magi |
| E-mail(s) |
albertomagi@gmail.com
|
| Organization name |
University of Florence
|
| Department |
Biochemical Sciences
|
| Street address |
Viale Morgagni 50
|
| City |
Firenze |
| ZIP/Postal code |
50134 |
| Country |
Italy |
| |
|
| Platform ID |
GPL1708 |
| Series (1) |
| GSE17596 |
Expression profiling of human mesoangioblasts indicates that S1P induces differentiation towards smooth muscle cells |
|
