|
| Status |
Public on Jul 31, 2017 |
| Title |
somatic cell from young donor [Y-SC] |
| Sample type |
RNA |
| |
|
| Source name |
somatic cell from young donor
|
| Organism |
Mus musculus |
| Characteristics |
cell type: somatic cell
age: young
|
| Treatment protocol |
Fibroblasts were collected from B6CBAF1 mouse E17.5 embryonic skin and 1.5-year-old tail tip skin. 10E6 fibroblasts were infected with retrovirus generated from pMX-mOCT4, pMX-mSOX2, pMX-mKLF4, and pEYK-mMYC in 6-well dishes with 0.5 ml of each viral supernatant (total 2 ml per well) and spun at 2500 rpm at RT for 90 min (BenchTop Centrifuge, BeckmanCoulter, Allegra-6R). For the generation of A-iPSC-ZSCAN10, the procedure was identical but in addition to the four reprogramming factors, we added a doxycycline-inducible system to overexpress ZSCAN10.”
|
| Growth protocol |
Mouse ESC and iPSC were cultured in ESC media containing 15% FBS and 1,000 U/ml of LIF (ESGRO® Leukemia Inhibitory Factor [LIF], 1 million units/1 mL). Doxycycline-inducible ZSCAN10 was induced in media supplemented with 2 μg/ml doxycycline (MP Biomedicals, doxycycline hyclate). Human iPSC were cultured in human ESC media (DMEM/F12, 20% knockout serum replacement, 10 ng/ml bFGF, and 0.1 mM -mercaptoethanol) on irradiated mouse embryonic feeders. Human and mouse fibroblasts were cultured in DMEM media with 10% FBS.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was reverse transcribed using the M-MuLV Reverse Transcriptase system (New England Biolabs), and the resulting cDNA was diluted. Synthesized cDNA solution was used for analysis.
|
| Label |
cy3
|
| Label protocol |
Biotinylated cRNA were prepared with the Ambion MessageAmp kit for Illumina arrays
|
| |
|
| Hybridization protocol |
Standard Illumina hybridization protocol
|
| Scan protocol |
Standard Illumina scanning protocol
|
| Data processing |
Data normalization was calculated with the Quantile normalization algorithm implemented in the BioConductor lumi package.
|
| |
|
| Submission date |
Jan 10, 2017 |
| Last update date |
Jul 31, 2017 |
| Contact name |
Hu Li |
| E-mail(s) |
li.hu@mayo.edu
|
| Organization name |
Mayo Clinic
|
| Department |
Molecular Pharmacology & Experimental Therapeutics
|
| Lab |
Systems Biology and Pharmacology
|
| Street address |
200 First Street, Gonda Building G19-408
|
| City |
Rochester |
| State/province |
MN |
| ZIP/Postal code |
55904 |
| Country |
USA |
| |
|
| Platform ID |
GPL6885 |
| Series (1) |
| GSE85365 |
Recovery of genomic stability by ZSCAN10 in induced pluripotent stem cells from aged donors |
|
