|
| Status |
Public on Sep 29, 2016 |
| Title |
Human Embyronic Stem Cell (ESCs) [H1-rep 2] |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
HESC-H1
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: H1
gender: Male
|
| Treatment protocol |
The cells were cultured in normal condition and were collected for microarray untill they were 70% confluency.
|
| Growth protocol |
Human ESCs and iPSCs were maintained on Matrigel (growth-factor-reduced; BD Biosciences)-coated plates with mTeSR1 (Stemcell Technologies) according to Wicell protocols. Cells were passaged every 4-5 days at ~80% confluence by using 0.02% EDTA (Versene). The NSCs were maintained on Poly-L-ornithine/ Laminin (Sigma) coated flasks with the N2B27 medium containing: DMEM/F12, N2 supplement (1:100), B27 supplement (1:50, Invitrogen), 100 mM nonessential amino acids (Invitrogen) plus 10 ng/mL FGF2 (R&D Systems), 10 ng/mL EGF (R&D Systems). The cells were passaged every 4-6 days at ~90% confluence by using Accutase. All NSCs used in this study were between passage 6-10.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA isolated was done using the ReliaPrep™ RNA Cell Minprep System (Promega);
|
| Label |
Cy3
|
| Label protocol |
The total RNA were labled with Cy3 (Samples) or Cy5 (References) by following the protocol of Two-Color Microarray-Based Gene Expression Analysis from Agilent.
|
| |
|
| Channel 2 |
| Source name |
Pooled Agilent's Universal Human Reference RNA
|
| Organism |
Homo sapiens |
| Characteristics |
cell lines: mixture of 10 human cell lines
reference: Pooled Agilent's Universal Human Reference RNA, catalog#740000
|
| Treatment protocol |
The cells were cultured in normal condition and were collected for microarray untill they were 70% confluency.
|
| Growth protocol |
Human ESCs and iPSCs were maintained on Matrigel (growth-factor-reduced; BD Biosciences)-coated plates with mTeSR1 (Stemcell Technologies) according to Wicell protocols. Cells were passaged every 4-5 days at ~80% confluence by using 0.02% EDTA (Versene). The NSCs were maintained on Poly-L-ornithine/ Laminin (Sigma) coated flasks with the N2B27 medium containing: DMEM/F12, N2 supplement (1:100), B27 supplement (1:50, Invitrogen), 100 mM nonessential amino acids (Invitrogen) plus 10 ng/mL FGF2 (R&D Systems), 10 ng/mL EGF (R&D Systems). The cells were passaged every 4-6 days at ~90% confluence by using Accutase. All NSCs used in this study were between passage 6-10.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA isolated was done using the ReliaPrep™ RNA Cell Minprep System (Promega);
|
| Label |
Cy5
|
| Label protocol |
The total RNA were labled with Cy3 (Samples) or Cy5 (References) by following the protocol of Two-Color Microarray-Based Gene Expression Analysis from Agilent.
|
| |
|
| |
| Hybridization protocol |
Oligoarray control targets and hybridization buffer (Agilent Gene Expression Hybridization Kit) were added, and samples were applied to microarrays enclosed in Agilent Microarray Hybridization Chambers. After hybridization, slides were washed sequential
|
| Scan protocol |
Scanned on a Agilent C Scanner
|
| Description |
Biological replicate 2 of 2. Human ESCs- H1.
|
| Data processing |
Agilent Feature Extraction (FE) software was used for background subtraction and LOWESS normalization.
|
| |
|
| Submission date |
Mar 26, 2015 |
| Last update date |
Apr 23, 2018 |
| Contact name |
LILI ZHU |
| E-mail(s) |
[email protected]
|
| Phone |
+441912418817
|
| Organization name |
Newcastle University
|
| Street address |
Central Parkway
|
| City |
Newcastle upon Tyne |
| State/province |
---- |
| ZIP/Postal code |
NE1 3 BZ |
| Country |
United Kingdom |
| |
|
| Platform ID |
GPL17077 |
| Series (1) |
| GSE67325 |
Human ESCs vs Human iPSCs & Human NSCs-ES vs Human NSCs-iPS |
|
| Relations |
| Reanalyzed by |
GSE113533 |
