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| Status |
Public on Oct 02, 2012 |
| Title |
Differentiation stage-specific donor memory in induced pluripotent stem cells (iPSC) generated from hepatic lineage cells |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
Recent studies suggested that embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) may represent different pluripotent states as defined by gene expression profiles and differentiation potential. Here we addressed a contribution of a lineage stage-specific donor cell memory in modulating the functional properties of iPSCs. iPSCs were generated from hepatic lineage cells at an early (hepatoblast-derived, HB-iPSCs) and end stage (adult hepatocyte, AH-iPSCs) of hepatocyte differentiation as well as from mouse fetal fibroblasts (MEF-iPSCs) using a lentiviral vector encoding four pluripotency-inducing factors Oct4, Sox2, Klf4, and c-Myc. All resulting iPS cell lines acquired iPSCs phenotype as judged by the accepted criteria including morphology, expression of pluripotency markers, silencing of transducing factors, capacity of multilineage differentiation in teratoma assay and normal diploid karyotype. However, hepatoblasts were more susceptible to reprogramming than either AH or MEF, and HB-iPSCs were more efficient in directed differentiation towards hepatocytic lineage as compared to AH-iPSCs, MEF-iPSCs or mESCs. Extensive comparative transcriptome analyses of the early passage iPSCs, donor cells and mESCs revealed that despite global similarities in gene expression patterns between generated iPSCs and mESCs, HB-iPSCs retained a transcriptional memory (7 up- and 20 down-regulated genes) typical of the original cells. Continuous passaging of HB-iPSCs abolished most of these differences including a superior capacity of hepatic re-differentiation. These results suggest that retention of lineage stage-specific donor memory in iPSCs may facilitate differentiation into donor cell type. The identified gene set may be helpful to improve hepatic differentiation for therapeutic application in liver disease modeling.
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| Overall design |
A total of 200 ng RNA from four independent biological replicates of MACS-sorted mESC and iPSC were linearly amplified according to manufactures’ specification (Ambion, Austin, Tx,). For in vitro transcription (IVT), reactions were incubated for 16 h at 37ºC. The efficiency of the single round amplification was measured by NanoDrop (ND1000, Thermo Scientific). Hybridization, washing, detection (Cy3-streptavidin, Amersham Biosciences, GE Healthcare), and scanning were performed on an illumina iScan system (Illumina) using reagents and following protocols supplied by the manufacturer. The biotinylated cRNA (750 ng/sample) was hybridized on Sentrix beadchips human Ref-8v3 for 18 h at 58ºC while rocking (5 rpm).
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| Contributor(s) |
Seo D, Lee S, Choi D, Marquardt JU, Thorgeirsson SS |
| Citation(s) |
22378611 |
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| Submission date |
Oct 20, 2011 |
| Last update date |
Jun 14, 2018 |
| Contact name |
Daekwan Seo |
| E-mail(s) |
[email protected]
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| Phone |
301-496-5688
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| Fax |
301-496-0734
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| Organization name |
NIH
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| Department |
NCI
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| Lab |
LEC
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| Street address |
37 Convent Dr. Rm 4140
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| City |
Bethesda |
| State/province |
MD |
| ZIP/Postal code |
20878 |
| Country |
USA |
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| Platforms (1) |
| GPL6885 |
Illumina MouseRef-8 v2.0 expression beadchip |
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| Samples (28)
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| Relations |
| BioProject |
PRJNA149353 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE33110_RAW.tar |
3.1 Mb |
(http)(custom) |
TAR |
| GSE33110_non-normalized.txt.gz |
5.5 Mb |
(ftp)(http) |
TXT |
| Processed data included within Sample table |
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