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| Status |
Public on Jun 30, 2014 |
| Title |
Differentiation of human limbal-derived induced pluripotent stem cells into limbal-like epithelium |
| Organism |
Homo sapiens |
| Experiment type |
Methylation profiling by array
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| Summary |
Limbal epithelial stem cell (LESC) deficiency represents a significant clinical problem especially in bilateral cases. Induced pluripotent stem cells (iPSC) may be a promising source of LESC, allowing standardized and continual propagation and banking. The objective of this study was to generate iPSC from human limbal epithelial cultures and differentiate them back into limbal epithelial cells using substrata mimicking the natural LESC niche. Using Yamanaka’s episomal vectors limbal-derived iPSC were reprogrammed from LESC cultured from donor corneoscleral rims and from human skin fibroblasts. A clone from limbal-derived iPSC expressed stemness markers, had a diploid karyotype, and produced teratomas in nude mice representing three germ layers. Compared to parental LESC, this clone had fewer specific gene methylation changes revealed using the Illumina Infinium Methylation 450k Beadchips than compared to skin fibroblasts. The expression of putative LESC markers was examined by quantitative RT-PCR and immunostaining in limbal-derived and fibroblast-derived iPSC cultured on denuded human amniotic membrane or denuded cornea. Limbal-derived iPSC had markedly stronger expression of PAX6, ABCG2, Np63, keratins 14, 15, 17, and N-cadherin than fibroblast-derived iPSC. On denuded corneas, limbal-derived iPSC showed the expression of differentiated corneal keratins 3 and 12. The data suggest that iPSC differentiation to a desired lineage may be facilitated by their generation from the same tissue. This may be related to preservation of parental tissue epigenetic methylation signatures in iPSC and use of biological substrata similar to the natural niche of parental cells. The data pave the way for generating transplantable LESC from limbal-derived iPSC.
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| Overall design |
Bisulphite converted DNA from the 12 samples were hybridised to the Illumina Infinium 450k Human Methylation Beadchip
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| Contributor(s) |
Sareen D, Saghizadeh M, Tang J, Funari VA, Punj V, Svendsen CN, Ljubimov AV |
| Citation(s) |
25069777 |
| Submission date |
Jan 08, 2014 |
| Last update date |
Mar 22, 2019 |
| Contact name |
Jie Tang |
| E-mail(s) |
jie.tang@cshs.org
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| Organization name |
Cedars Sinai Medical Center
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| Street address |
8700 Beverly Blvd
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| City |
Los Angeles |
| ZIP/Postal code |
90048 |
| Country |
USA |
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| Platforms (1) |
| GPL13534 |
Illumina HumanMethylation450 BeadChip (HumanMethylation450_15017482) |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA233965 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE53918_RAW.tar |
183.1 Mb |
(http)(custom) |
TAR |
| GSE53918_signal_iintensities.txt.gz |
69.8 Mb |
(ftp)(http) |
TXT |
| Processed data included within Sample table |
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