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| Status |
Public on Dec 31, 2010 |
| Title |
Sox2 Levels Define Pluripotent Ground States and Reprogramming Potential |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
Pluripotent stem cell lines derived from embryos of different stages have distinct pluripotent ground states, but similar levels of the transcription factor Oct4. Epiblast-derived pluripotent stem cells (EpiSCs), in contrast to embryonic stem (ES) cells, cannot form chimeras. We show that EpiSCs express lower levels of the transcription factors Sox2 and Klf4 than ES cells and have limited reprogramming potential, as shown by cell fusion. Sox2 overexpression dramatically increases the reprogramming potential, chimera formation, and germline contribution of EpiSCs. Therefore, although Oct4 is essential for reprogramming, the level of Sox2 defines both the reprogramming capability and the pluripotent ground states.
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| Overall design |
RNA samples to be analyzed on microarrays were prepared using Qiagen RNeasy columns with on-column DNA digestion. 300 ng of total RNA per sample was used as input into a linear amplification protocol (Ambion), which involved synthesis of T7-linked double-stranded cDNA and 12 hrs of in-vitro transcription incorporating biotin-labelled nucleotides. Purified and labelled cRNA was then hybridized for 18 hrs onto MouseRef-8 v2 expression BeadChips (Illumina) according to the manufacturer's instructions. After washing, as recommended, chips were stained with streptavidin-Cy3 (GE Healthcare) and scanned using the iScan reader (Illumina) and accompanying software. Samples were hybridized as biological replicates. 12 sample types were analyzed, each of them in duplicate.
ESCm: Mouse ESC male; ESCf: Mouse ESC OG2 female; F9 EC: F9 EC (mouse embryonic carcinoma cell); F9-Sox2: F9 EC (mouse embryonic carcinoma cell) overexpressing wild type Sox2; EpiSCf: Mouse EpiSC OG2 female; Epi-Sox2f: Mouse EpiSC Sox2 (OG2 female) overexpressing wild type Sox2; P19 EC: P19 EC (mouse embryonic carcinoma cell); P19-Sox2: P19 EC (mouse embryonic carcinoma cell) overexpressing wild type Sox2; EpiSCm: Mouse EpiSC (GOF18 male) (duplicates); EpiSox2mL2: Mouse EpiSC Sox2 (GOF18 male) overexpressing wild type Sox2 cultured in condition EpiSC medium (CM); EpiSox2mE1: Mouse EpiSC Sox2 (GOF18 male) overexpressing wild type Sox2 cultured in ESC medium (ESC like1); EpiSox2mE2: Mouse EpiSC Sox2 (GOF18 male) overexpressing wild type Sox2 cultured in ESC medium (ESC like2).
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| Contributor(s) |
Han DW, Do JT, Tapia N, Joo JY, Araúzo-Bravo MJ, Bernemann C, Greber B, Stehling M, Sterneckert J, Zaehres H, Schöler HR |
| Citation missing |
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| Submission date |
Mar 23, 2009 |
| Last update date |
Jan 15, 2022 |
| Contact name |
Marcos J. Araúzo-Bravo |
| E-mail(s) |
mararabra@yahoo.co.uk
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| Phone |
+34 943 00 6108
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| Organization name |
Max Planck Institute for Molecular Biomedicine
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| Department |
Cell and Developmental Biology
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| Lab |
Computational Biology and Bionformatics
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| Street address |
Rogentstrasse
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| City |
Muenster |
| ZIP/Postal code |
48149 |
| Country |
Germany |
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| Platforms (1) |
| GPL6885 |
Illumina MouseRef-8 v2.0 expression beadchip |
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| Samples (24)
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| GSM385571 |
ESCf, rep2 |
| GSM385572 |
F9 EC, rep1 |
| GSM385573 |
F9 EC, rep2 |
| GSM385574 |
F9-Sox2, rep1 |
| GSM385575 |
F9-Sox2, rep2 |
| GSM385576 |
EpiSCf, rep1 |
| GSM385577 |
EpiSCf, rep2 |
| GSM385578 |
Epi-Sox2f, rep1 |
| GSM385579 |
Epi-Sox2f, rep2 |
| GSM385580 |
P19 EC, rep1 |
| GSM385581 |
P19 EC, rep2 |
| GSM385582 |
P19-Sox2, rep1 |
| GSM385583 |
P19-Sox2, rep2 |
| GSM385584 |
EpiSCm, rep1 |
| GSM385585 |
EpiSCm, rep2 |
| GSM385586 |
EpiSox2mL2, rep1 |
| GSM385587 |
EpiSox2mL2, rep2 |
| GSM385588 |
EpiSox2mE1, rep1 |
| GSM385589 |
EpiSox2mE1, rep2 |
| GSM385590 |
EpiSox2mE2, rep1 |
| GSM385591 |
EpiSox2mE2, rep2 |
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| Relations |
| BioProject |
PRJNA115777 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE15358_RAW.tar |
3.1 Mb |
(http)(custom) |
TAR |
| GSE15358_non-normalized.txt |
12.0 Mb |
(ftp)(http) |
TXT |
| Processed data included within Sample table |
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