|
| Status |
Public on Oct 17, 2012 |
| Title |
Undifferentiated hESCs_Rep1 |
| Sample type |
RNA |
| |
|
| Source name |
hESCs treated with FGF2 (20 ng/ml)
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: hESC
cell line: H9
treatment: FGF2 (20 ng/ml)
|
| Treatment protocol |
The cells were passaged and maintained in MEF-conditioned medium (Xu et al., 2001) till they reached a confluency of ~40-50%. They were rinsed with PBS, followed by the various treatments under defined conditions (N2B27) (Yao et al., 2006): BMP4 (10 ng/ml) for 3 Hours and 5 Days; BMP4 (10 ng/ml) + SB431542 (20 µM) for 3 Hours, 1 Day, 3 Days and 5 Days; BMP4 (10 ng/ml) + SB431542 (20 µM) + SU5402 (20 µM) for 3 Hours, 1 Day, 3 Days and 5 Days; BMP4 (10 ng/ml) + SB431542 (20 µM) + FGF2 (20 ng/ml) for 3 Hours, 1 Day, 3 Days and 5 Days; SU5402 (20 µM) for 1 Day, 3 Days and 5 Days
|
| Growth protocol |
The experiments were carried out using the Human Embryonic Stem Cell line H9 obtained from WiCell and cultured on matrigel-coated plates. The cells were passaged and maintained in MEF-conditioned medium (Xu et al., 2001) till they reached a confluency of ~40-50%. They were rinsed with PBS, followed by the various treatments under defined conditions (N2B27) (Yao et al., 2006).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA isolations were performed using the RNeasy Mini Kit (Qiagen). The cells were directly lysed with lysis buffer in the cell-culture wells, after rinsing with PBS. Total RNA from human Placenta was purchased from Clontech (636527).
|
| Label |
Biotin
|
| Label protocol |
500 ng DNAse-free total RNA was biotin-labelled to generate cRNA employing a linear amplification kit (Ambion).
|
| |
|
| Hybridization protocol |
The labelled cRNA samples were hybridised onto HumanRef-8 v3 chips following the manufacturer’s instructions (Ilumina).
|
| Scan protocol |
The hybridized chips were scanned following the manufacturer’s instructions (Ilumina).
|
| Description |
Biological Replicate 1
|
| Data processing |
Rank invariant normalization method from BeadStudio 3.0 software (Illumina) was used for data normalization.
|
| |
|
| Submission date |
Jun 21, 2011 |
| Last update date |
Oct 17, 2012 |
| Contact name |
smita sudheer |
| E-mail(s) |
sudheer@molgen.mpg.de
|
| Organization name |
Max Planck Institute for Molecular Genetics
|
| Street address |
Ihnestraße 63-73
|
| City |
Berlin |
| ZIP/Postal code |
14195 |
| Country |
Germany |
| |
|
| Platform ID |
GPL6883 |
| Series (1) |
| GSE30125 |
FGF inhibition directs BMP4-mediated differentiation of Human Embryonic Stem Cells to syncytiotrophoblast |
|
