|
| Status |
Public on Apr 30, 2009 |
| Title |
hMSCs_BV-transduced_rep4 |
| Sample type |
RNA |
| |
|
| Source name |
hMSCs, BV-transduced
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: Human mesenchymal stem cells (hMSCs)
hmscs passage number: 11th passage
|
| Treatment protocol |
The virus transduction was performed on 6-well plates. Depending on the MOI, a certain volume of virus supernatant was mixed with NaHCO3-deficient a-MEM medium to adjust the final volume to 500 ml (per well). The transduction was initiated by adding the virus mixture to the cells and continued by gently shaking the plates on a rocking plate for 4 h at 25-27 oC. For mock-transduction, the cells were incubated under the same conditions with the solution consisting of 400 ml NaHCO3-deficient a-MEM and 100 ml fresh TNM-FH medium.
|
| Growth protocol |
Bone marrow-derived hMSCs were obtained from Cambrex Co., selected, enriched, cultured in a-MEM medium containing 10% fetal bovine serum (FBS, Hyclone) , and expanded to passage 11 for all experiments.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted with the RNeasy® Mini Kit (Qiagen) according to manufacturer's specification.
|
| Label |
Cy5
|
| Label protocol |
cDNA were synthesized from the extracted total RNA by reverse transcription followed by second strand synthesis using the Amino Allyl MessageAmpII aRNA amplification Kit (Ambion) according to the manufacturer's specification. Amino allyl modified aRNA produced was subsequently coupled to Cy5 dye label.
|
| |
|
| Hybridization protocol |
Cy5-labeled aRNA were suspended in OneArray™ hybridization buffer (provided in HOA product package) at a final volume of 180 ul per bridization. The pre-hybridization blocking, array hybridization, and post-hybridization washes were performed according to the instruction provided in the HOA User Guide.
|
| Scan protocol |
The arrays were scanned using DNA Microarray Scanner (Agilent Technologies) and the fluorescent intensities are extracted from the generated images by following the instructions and the conditions described in HOA User Guide.
|
| Description |
hMSCs_BV-transduced_rep4
|
| Data processing |
The raw data was preprocessed by log2 transformation and Global Lowess normalization. The preprocessed data was then analyzed by Limma package of R software. In this study, the significantly changed genes were identified and filtered based on one-way ANOVA (adjusted p<0.05).
|
| |
|
| Submission date |
Apr 24, 2009 |
| Last update date |
Jun 26, 2012 |
| Contact name |
Yu-Chen Hu |
| E-mail(s) |
yuchen@che.nthu.edu.tw
|
| Organization name |
National Tsing Hua University
|
| Department |
Chemical Engineering
|
| Street address |
101, Section 2, Kuang-Fu Road
|
| City |
Hsinchu |
| ZIP/Postal code |
30011 |
| Country |
Taiwan |
| |
|
| Platform ID |
GPL6254 |
| Series (1) |
| GSE15810 |
The global responses of human bone marrow-derived mesenchymal stem cells to baculovirus transduction |
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