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Generation and Characterization of Stable Pig Pre-gastrulation Epiblast Stem Cell Lines [ChIP-seq]

ABSTRACT: Generation and Characterization of Stable Pig Pre-gastrulation Epiblast Stem Cell Lines [ChIP-seq]

PROVIDER: PRJNA706843 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR13833769_1.fastq.gz	Fastqsanger.gz
	SRR13833769_2.fastq.gz	Fastqsanger.gz
	SRR13833770_1.fastq.gz	Fastqsanger.gz
	SRR13833770_2.fastq.gz	Fastqsanger.gz
	SRR13833771_1.fastq.gz	Fastqsanger.gz

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Generation and Characterization of Stable Pig Pre-gastrulation Epiblast Stem Cell Lines

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Generation and Characterization of Stable Pig Pre-gastrulation Epiblast Stem Cell Lines [HiC]

Project description:Despite ongoing attempts since the 1990s, no stable embryonic stem cell line has been established in pigs. Here, guided by insights from large-scale single-cell RNA-sequencing profiling of pig embryos from embryonic day (E) 0 to E14, we developed an in vitro culture system for establishing and maintaining stable pluripotent stem cell lines from pig E10 pre-gastrulation epiblasts (pgEpiSCs). Enabled by chemical inhibition of WNT-related signaling in culture, the pgEpiSCs maintain their pluripotency and normal karyotypes after more than 200 passages. Strikingly, ultra-deep in situ Hi-C analysis revealed functional impacts of 3D-spatial associations on the transcriptional regulation of pluripotency marker genes in the pgEpiSCs. Practically, we confirmed that the pgEpiSCs can readily tolerate at least three rounds of successive gene editing and generated cloned gene-edited live piglets. Our findings deliver on the long-anticipated promise of large model animal stem cells and open new avenues for biological research, animal husbandry, and biomedicine.

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Project description:Genome integration-free pig induced pluripotent stem cells (iPSCs) bring tremendous value in preclinical testing of regenerative medicine, as well as conservation and exploitation of endangered or rare local pig idioplasmatic resources. However, due to a lack of appropriate culture medium, efficient induction and stable maintenance of pig iPSCs with practical value remains challenging. Here, we established an efficient induction system for exogenous gene-independent iPSCs under WNT-inhibited condition previously used for generation of stable pig pre-gastrulation epiblast stem cell lines (pgEpiSCs). WNT suppression was found to play an essential role in establishment of exogenous gene-independent iPSCs. Strikingly, stable integration-free pig iPSCs could be reprogrammed from pig somatic cells using episomal vectors in this cultured condition. The iPSCs had pluripotency features and transcriptome characteristics approximating pgEpiSCs. More importantly, this induction system may be used to generate integration-free iPSCs from elderly disabled rare local pig somatic cells, and the iPSCs could be gene-edited and used as donor cells for nuclear transfer. Our results provide novel insights into potential applications for genetic breeding of livestock species and pre-clinical evaluation of regenerative medicine.

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