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| Status |
Public on Nov 09, 2023 |
| Title |
Efficient derivation of transgene-free porcine induced pluripotent stem cells enables in vitro modeling of species-specific developmental timing |
| Organism |
Sus scrofa |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Sus domesticus (pig) are an excellent large mammalian model for comparative studiesdue to their relatively comparable physiology and organ size to humans. The derivation of transgene-free porcine pluripotent stem cells (PiPSCs) will therefore benefit the development of porcine-specific models for regenerative biology and its medical applications. In the past, this effort has been hampered by a lack of understanding of the signaling milieu that stabilizes the porcine pluripotent state in vitro. Here, we report that transgene-free PiPSCs can be efficiently derived from porcine fibroblasts by episomal vectors along with microRNA-302/367 using optimized protocols tailored for this species. Established PiPSCs can robustly differentiate into derivates representing the primary germ layers in vitro and in vivo. Furthermore, the transgene- free PiPSCs preserve intrinsic species-specific developmental timing in culture. This capacity is demonstrated by establishing a porcine in vitro segmentation clock model that, for the first time, displays a specific periodicity at ~ 3.7 hours, a time scale recapitulating in vivo porcine somitogenesis. We therefore propose that these transgene-free PiPSCs can serve as a powerful tool for modeling development and disease, informing both conserved and unique features of mammalian pluripotency and developmental timing mechanisms.
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| Overall design |
A total of 36 bulk RNA-seq samples: (1) compare undifferentiated clonal PiPSC lines to fibroblasts. (2) PiPSC differentiation to various lineage-specific progenitors. (3) PiPSC differentiation to primitive streak, PSM and somite cell fates. To construct the RNA-seq library, total RNA from each sample (an input of ~100 ng total (RNA) was used following the LM-seq protocol
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| Contributor(s) |
Chu L, Conrad JV, Meyer S, Mamott D, Duffin B, Ramesh PS, Rusteika M, Neira JA, Zhang J, Hiles E, Higgins E, Steill J, Stewart R, Thomson JA, Bautista M, Freeman J, Ni Z, Liu S, Ungrin M, Rancourt D, Clegg DO |
| Citation(s) |
37949072 |
| Submission date |
Mar 22, 2023 |
| Last update date |
Jan 02, 2024 |
| Contact name |
Li-Fang (Jack) Chu |
| E-mail(s) |
lifangjack.chu@ucalgary.ca
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| Organization name |
University of Calgary
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| Lab |
Chu Laboratory
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| Street address |
2500 University Drive NW
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| City |
Calgary |
| State/province |
Alberta |
| ZIP/Postal code |
T2N 1N4 |
| Country |
Canada |
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| Platforms (2) |
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| Samples (36)
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| Relations |
| BioProject |
PRJNA947624 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE227985_ec.tsv.gz |
1.3 Mb |
(ftp)(http) |
TSV |
| GSE227985_tpm.tsv.gz |
1.2 Mb |
(ftp)(http) |
TSV |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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