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LIF-Dependent, Pluripotent Stem Cells Established from Inner Cell Mass of Porcine Embryos

ABSTRACT: The pig is important for agriculture and as an animal model in human and veterinary medicine, yet, despite over 20 years of effort, it has proved a difficult species from which to generate pluripotent stem cells analogous to those derived from mouse embryos. Here we report the production of LIF-dependent, so called naïve type, pluripotent stem cells from the inner cell mass of porcine blastocysts by up-regulating expression of KLF4 and POU5F1. These cells resemble mouse ES cells and are distinct from the FGF2-dependent, induced pluripotent cell type derived from porcine somatic cells. High throughput SNP chip genotyping was conducted on Illumina's Porcine SNP60 BeadChip (WG-410, a service provided by Geneseek, NE, http://www.neogen.com/GeneSeek/). The results exhibited that the two lines pluripotent stem cells from the inner cell mass of porcine blastocysts were porcine origin and genetically distinct. Porcine pluripotent stem cells were derived from the inner cell mass with transduction with human KLF4 by lentiviral transduction.

ORGANISM(S): Sus scrofa

SUBMITTER: Bhanu Telugu

PROVIDER: E-GEOD-26436 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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LIF-Dependent, Pluripotent Stem Cells Established from Inner Cell Mass of Porcine Embryos

Project description:The pig is important for agriculture and as an animal model in human and veterinary medicine, yet, despite over 20 years of effort, it has proved a difficult species from which to generate pluripotent stem cells analogous to those derived from mouse embryos. Here we report the production of LIF-dependent, so called naïve type, pluripotent stem cells from the inner cell mass of porcine blastocysts by up-regulating expression of KLF4 and POU5F1. These cells resemble mouse ES cells and are distinct from the FGF2-dependent, induced pluripotent cell type derived from porcine somatic cells. High throughput SNP chip genotyping was conducted on Illumina's Porcine SNP60 BeadChip (WG-410, a service provided by Geneseek, NE, http://www.neogen.com/GeneSeek/). The results exhibited that the two lines pluripotent stem cells from the inner cell mass of porcine blastocysts were porcine origin and genetically distinct.

2011-01-05 | GSE26436 | GEO

LIF-Dependent, Pluripotent Stem Cells Established from Inner Cell Mass of Porcine Embryos

Project description:The pig is important for agriculture and as an animal model in human and veterinary medicine, yet, despite over 20 years of effort, it has proved a difficult species from which to generate pluripotent stem cells analogous to those derived from mouse embryos. Here we report the production of LIF-dependent, so called naM-CM-/ve type, pluripotent stem cells from the inner cell mass of porcine blastocysts by up-regulating expression of KLF4 and POU5F1. These cells resemble mouse ES cells and are distinct from the FGF2-dependent, induced pluripotent cell type derived from porcine somatic cells. Transcript profiling utilized Affymetrix porcine microarrays were conducted to compare the gene expressions associated with pluripotency in the two LIF-dependent pESK lines (pESK I & II, passage 14 & 5, respectively) with a naM-CM-/ve phenotype and two porcine iPSC lines (ID4 & ID6, both passage 7, GSE15472) with a primed phenotype that re-programming of porcine fetal fibroblasts (EGFP-PFF, GSE15472). The pESK lines clustered separately from the porcine iPSC lines, EGFP-PFF and primary cultures established from explants of porine umbilical cord (PUC). Porcine pluripotent stem cells were derived from the inner cell mass with transduction with human KLF4 by lentiviral transduction.

2011-12-02 | E-GEOD-26369 | biostudies-arrayexpress

LIF-Dependent, Pluripotent Stem Cells Established from Inner Cell Mass of Porcine Embryos

Project description:The pig is important for agriculture and as an animal model in human and veterinary medicine, yet, despite over 20 years of effort, it has proved a difficult species from which to generate pluripotent stem cells analogous to those derived from mouse embryos. Here we report the production of LIF-dependent, so called naïve type, pluripotent stem cells from the inner cell mass of porcine blastocysts by up-regulating expression of KLF4 and POU5F1. These cells resemble mouse ES cells and are distinct from the FGF2-dependent, induced pluripotent cell type derived from porcine somatic cells. Transcript profiling utilized Affymetrix porcine microarrays were conducted to compare the gene expressions associated with pluripotency in the two LIF-dependent pESK lines (pESK I & II, passage 14 & 5, respectively) with a naïve phenotype and two porcine iPSC lines (ID4 & ID6, both passage 7, GSE15472) with a primed phenotype that re-programming of porcine fetal fibroblasts (EGFP-PFF, GSE15472). The pESK lines clustered separately from the porcine iPSC lines, EGFP-PFF and primary cultures established from explants of porine umbilical cord (PUC).

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ChIP-Seq of STAT3 in 3iL Human Embryonic Stem Cells

Project description:Human embryonic stem cells (hESCs) are isolated from the inner cell mass of the blastocysts. The pluripotent properties of hESCs enable the derivation of cell-types or tissues of different lineages for potential applications such as therapeutics discovery and regenerative medicine. Even though hESCs are pluripotent, differences have been observed when compared to the native pluripotent epiblast cells of the blastocyst. We use a chemical approach (3iL: 3 small molecule inhibitor and cytokine) to induce an expression signature that more closely resembles native pluripotent cells. This experiment is STAT3 binding data in 3iL hESCs.

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Lineage specification and pluripotency revealed by transcriptome analysis from oocyte to blastocyst in pig

Project description:The inner cell mass in blastocyst is the origin of all the somatic and germ cells in mammals, and of pluripotent stem cells in vitro. As the conserved principles between pig and human, here we performed comprehensive single-cell RNA-seq for porcine early embryos from oocyte to early blastocyst. We show the specification of inner cell mass and trophectoderm in morula, and the molecular signature of the precursors. We demonstrate the existence of naïve pluripotency signature in morula and inner cell mass of early blastocyst, and the specific pluripotent genes and the activity of signalling pathways highlight the characteristics of the naïve pluripotency. We observe absence of dosage compensation with respect to X-chromosome in morula, and incomplete dosage compensation in early blastocyst. However, the dynamics of dosage compensation may be independent on the expression of XIST induced X-chromosome inactivation. Our study describes molecular landmarks of embryogenesis in pig that will provide a better strategy for derivation of porcine pluripotent stem cells and improve the research in regenerative medicine.

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IRF-1 expressed in the early blastocyst inner cell mass of pigs enhances the pluripotency of induced pluripotent stem cells

Project description:Naïve porcine stem cells is supposed to rely on the regulation of unique transcription factor in the early inner cell mass. The expression of Interferon regulatory factor 1 (IRF-1) were detected in aggregation in d6~7 somatic cell nuclear transfer blastocysts, which was supposed to regulate the pluripotency of naïve state. But IRF-1 is reported to be involved in the response to viral infections and initiate a rapid proinflammatory response, while the function of IRF-1 on pluripotency need to be explored. Then the heterogenicity of IRF-1 was associated with pluripotency in pig induced pluripotent cells and overexpression of IRF-1 promoted the gene expression profile of pluripotent genes as would be expected when cells approach the naïve state

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RNA-Seq of 3iL human embryonic stem cells

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Sus scrofa

Project description:LIF-Dependent, Pluripotent Stem Cells Established from Inner Cell Mass of Porcine Embryos

| PRJNA136815 | ENA