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| Status |
Public on Dec 03, 2010 |
| Title |
Reprogramming of human primary somatic cells by OCT4 and chemical compounds |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Human induced pluripotent stem cells (hiPSCs) can be practically derived from neonatal human epidermal keratinocytes (NHEKs) transduced with OCT4, by a novel cocktail of small molecules including 5 μM PS48, 0.25 mM NaB, 0.5 μM A-83-01 and 0.5 μM PD0325901. These iPSCs are morphologically, molecularly and functionally similar to pluripotent hESCs.
NHEKs were cultured in a 100 mm tissue culture dish and transduced 3 times (3-4 hours each transduction) with freshly produced lentivirus supernatants. 1,000,000 transduced NHEKs were seeded on the irradiated x-ray inactivated CF1 MEF feeder cells in a 100-mm dish and cultured in KCM and treated with 5 μM PS48, 0.25 mM NaB (Stemgent) and 0.5 μM A-83-01 (Stemgent) for 2 weeks, followed by changing half volume of media to hESCM and supplementing with 5 μM PS48, 0.25 mM NaB and 0.5 μM A-83-01 for another 2 weeks. Then cell culture media were changed to hESCM and supplemented with 5 μM PS48, 0.25 mM NaB, 0.5 μM A-83-01 and 0.5 μM PD0325901 (Stemgent) for additional four weeks.The culture was split by Accutase (Millipore) and treated with 1 μM Thiazovivin (Stemgent) in the first day after splitting. The iPSC colonies stained positive by Alexa Fluor 555 Mouse anti-Human TRA-1-81 antibody (BD Pharmingen) were picked up for expansion on feeder cells in hESCM and cultured routinely.
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| Overall design |
Global gene expression analysis of hiPSCs
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| Contributor(s) |
Zhu S, Ding S |
| Citation(s) |
21112560 |
| Submission date |
Nov 09, 2010 |
| Last update date |
Mar 20, 2017 |
| Contact name |
Saiyong Zhu |
| E-mail(s) |
zhusy@scripps.edu
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| Organization name |
The Scripps Research Institute
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| Street address |
10550 North Torrey Pines Road,
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| City |
La Jolla |
| State/province |
CA |
| ZIP/Postal code |
92037 |
| Country |
USA |
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| Platforms (1) |
| GPL6883 |
Illumina HumanRef-8 v3.0 expression beadchip |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA134453 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE25218_RAW.tar |
3.9 Mb |
(http)(custom) |
TAR |
| GSE25218_non-normalized.txt.gz |
1.1 Mb |
(ftp)(http) |
TXT |
| Processed data included within Sample table |
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