Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Derivation of authentic porcine embryonic stem cells using defined culture conditions

ABSTRACT: Pig embryonic stem cells (ESCs) have been considered as an important candidate for preclinical researches on human therapy. However, the lack of understanding of pig pluripotent networks has hampered establishment of authentic pig ESCs. Here, we report that FGF2, ACTVIN, and WNT signaling is essential for maintaining pig pluripotency in vitro. Pig ESC lines derived by stimulating three signlaings formed colonies of flattened monolayer morphology. Newly derived ESCs were stably maintained over an extended period, and were capable of forming teratomas comprising three germ layers. Immunostaining showed that the stem cells expressed pluripotency markers including OCT4, SOX2, NANOG, SSEA1, and SSEA4. Transcriptome analysis showed that pig ESCs were developmentally similar to late epiblasts of preimplantation embryos and in terms of biological functions resembled human rather than mouse ESCs. However, the pig ESCs had distinct features such as coexpression of SSEA1 and SSEA4, two active X chromosomes, and a unique transcriptional pattern. In conclusion, we successfully derived authentic pig ESCs using novel cell culture conditions. Our findings will facilitate both the development of large animal models for human stem cell therapy and the generation of pluripotent stem cells from other domestic animals for agricultural use.

ORGANISM(S): Sus scrofa

PROVIDER: GSE120031 | GEO | 2019/07/08

REPOSITORIES: GEO

ACCESS DATA

Json Xml

Similar Datasets

Molecular and functional features demarcate embryonic and induced pluripotent stem cells in rabbits

Project description:Little is known about the molecular and functional features of pluripotent stem cells (PSCs) in rabbits. To address this, we derived and characterized two types of rabbit PSCs from the same breed of New Zealand White rabbits: four lines of embryonic stem cells (rbESCs), and three lines of induced PSCs (rbiPSCs) that resulted from reprogramming adult skin fibroblasts. All lines were dependent on fibroblast growth factor 2. All rbESC lines exhibited molecular and functional properties typically associated with primed pluripotency. The rbESC lines also exhibited a cell cycle with a prolonged G1 phase and a DNA damage checkpoint prior to entry into the S phase, which are two features typically associated with the somatic cell cycle. In contrast, the rbiPSC lines exhibited some characteristics of naÃ¯ve pluripotency as defined in rodents, including resistance to single cell dissociation by trypsin, robust activity of the distal enhancer of the mouse Oct4 gene, and expression of naÃ¯ve-specific genes. According to gene expression profiles, rbiPSCs were closer to the rabbit inner cell mass than rbESCs. We propose that rbiPSCs self-renew in an intermediate state between naÃ¯ve and primed pluripotency, which represents a key step toward the generation of bona fide naÃ¯ve PSC lines in rabbits The patterns of stage specific embryonic antigen (SSEA) expression were also assessed between cell lines. All two rbiPSC lines exhibited heterogeneous expression of SSEA1 and SSEA4 that leads to obtain 3 different cell sub-populations (SSEA1+, SSEA1+/4+, SSEA1-/4-). All four rbESC lines expressed only SSEA1 and not SSEA4. Then, for this category of cell lines, we used 3 different cell sub-populations (SSEA1+, SSEA1-, and total which contained all cell types). Three independent experiments were performed for each line and for each cell sub-population.

2013-01-11 | E-GEOD-43403 | biostudies-arrayexpress

Expression data from ESC and in vitro derived somatic cells and germ cells

Project description:We performed gene expression profiling on in vitro derived PGCs, undifferentiated ESCs, and somatic cells from the EB to examine germ cell expression in ESC-derived cells All cells were collected using fluorescence activated cell sorting to isolate SSEA1+/cKit+ ESCs, SSEA1+/cKitbright PGCs, Oct4-gfp+/cKitbright PGCs, and SSEA1-/cKit- somatic cells and Oct4-gfp-/cKit- somatic cells.

2011-12-15 | E-GEOD-33121 | biostudies-arrayexpress

Identification of differentiation status of human embryonic stem cell cultures.

Project description:Two stages of culture were tested. ESC is the pluripotent stage expressing SSEA4 and Oct4 but not SSEA1. NSC is a neural-committed stage expressing Nestin but not beta-tubulin. Three cell lines were tested: H1, HSF1, and HSF6.

2009-06-01 | GSE15206 | GEO

Tet-mediated imprinting erasure following reprogramming of spermatogonial stem cells to induced pluripotent stem cells

Project description:We sequenced mRNA from three preparations each of murine Gfra1+ spermatogonial stem cells (SSC), SSC- derived SSEA1+ induced pluripotent stem cells (iPSC), and fibroblast (skin derived mesenchymal cells) derived SSEA1+ iPSC

2015-01-12 | GSE64856 | GEO

Derivation of formative-like pluripotent stem cells from mammalian embryos [ChIP-Seq]

Project description:Two phases of pluripotency, naïve and primed, have been captured in vitro and studied in details1. A third formative phase was recently proposed to exist between naïve and primed phases2. Formative pluripotency entails permissiveness for direct primordial germ cell (PGC) induction and competence for blastocyst chimeras, and is characterized by transcriptional and epigenetic features intermediate of naïve and primed pluripotency. To date, however, stable pluripotent stem cells (PSCs) harboring formative features haven’t been derived from early mammalian embryos. Here we develop a method which enabled the derivation and culture of stable formative-like embryonic stem cells (ESCs) from mouse blastocysts. Formative-like mouse ESCs share molecular features characteristic of early post-implantation epiblasts and are competent for PGC-like cell induction and blastocyst chimera formation. The same culture also supported the derivation of ESCs and transgene-free induced pluripotent stem cells (iPSCs) from horse blastocysts and fibroblasts, respectively. Horse ESCs/iPSCs transcriptionally resembled mouse formative cells, and could also be directly induced into PGC-like cells. Formative-like horse iPSCs could efficiently chimerize horse, mouse, goat, sheep and pig embryos. Stable formative-like PSCs will be invaluable for studying mammalian pluripotency, and our method may be broadly applicable for the derivation of PGC and chimera competent PSCs from other mammalian species.

2020-11-27 | GSE135990 | GEO

Derivation of formative-like pluripotent stem cells from mammalian embryos [RNA-Seq]

2020-11-27 | GSE135989 | GEO

OCT4 IP-MS in Naive and Primed Human ESCs

Project description:Understanding the molecular underpinnings of pluripotency is a prerequisite for optimal maintenance and application of embryonic stem cells (ESCs). While the protein-protein interactions of core pluripotency factors have been identified in mouse ESCs, their interactome in human ESCs (hESCs) has not to date been explored. Here we mapped the OCT4 interactomes in naïve and primed hESCs, revealing extensive connections to mammalian ATP-dependent nucleosome remodeling complexes.

2021-09-12 | PXD026556 | Pride

A genome-wide RNAi screen identifies genes and networks governing hematopoietic stem cell development [Illumina_Expression]

Project description:We report a genome-wide microarray analysis of gene expression in mouse embryonic stem cell (ESC) and in vitro ES-derived cells. iHoxB4 ESCs were transduced with a library of shRNAs targeting >15,000 genes, and were differentiated towards hematopoietic stem and progenitor cells by forcing HoxB4 expression in the cells. Five cell populations were isolated on differentiation days 6 and 20. Differentiated mesodermal/endothelium cell population D6F (Ssea1-Flkl+Cxcr4-), endodermal cell population D6C (Ssea1-Flkl-Cxcr4+), D20LS (Lin-Sca1+c-Kit–), D20LK (Lin–Sca1–c-Kit+), and D20LSK (Lin–Sca1+c-Kit+) cells were purified by FACS. Transduced iHoxB4 cells were analyzed as undifferentiated control cells. The experiment was performed three times independently.

2016-12-31 | GSE86853 | GEO

eIF4A3 interactome by IP-MS in mouse ESCs

Project description:RNA helicases are involved in multiple steps of RNA metabolism to direct their roles in gene expression, yet their functions in pluripotency control remain largely unexplored. Starting from an RNAi screen of RNA helicases, we identified that eIF4A3, a DEAD-box (Ddx) helicase component of the exon junction complex (EJC), is essential for the maintenance of embryonic stem cells (ESCs). We mapped the eIF4A3 interactomes in mouse ESCs, revealing that eIF4A3 is widely involved in the post-transcriptional regulation of gene expression. Mechanistically, we show that eIF4A3 post-transcriptionally controls the pluripotency-related cell cycle regulators and that its depletion causes cellular differentiation via cell cycle dysregulation. Specifically, eIF4A3 is required for the efficient nuclear export of Ccnb1 mRNA, which encodes Cyclin B1, a key component of the pluripotency-promoting pathway during cell cycle progression of ESCs. Our results reveal a previously unappreciated role of eIF4A3 and its associated EJC in the post-transcriptional cell cycle control in maintaining stem cell pluripotency.

2023-03-11 | PXD031362 | Pride

miR-371-3 expression predicts neural differentiation potential in human pluripotent stem cells

Project description:The use of pluripotent stem cells in regenerative medicine and disease modeling is complicated by the variation in differentiation properties between lines. In this study, we characterized 13 human embryonic stem cell. (hESC) and 26 human induced pluripotent stem cell (hiPSC) lines to identify markers that predict neural differentiation behavior. At a general level, markers previously known to distinguish mouse ESCs from epiblast stem cells (EpiSCs) correlated with neural differentiation behavior. More specifically, quantitative analysis of miR-371-3 expression prospectively identified hESC and hiPSC lines with differential neurogenic differentiation propensity and in vivo dopamine neuron engraftment potential. Transient KLF4 transduction increased miR-371-3 expression and altered neurogenic behavior and pluripotency marker expression. Conversely, suppression of miR- 371-3 expression in KLF4-transduced cells rescued neural differentiation propensity. miR-371-3 expression level therefore appears to have both a predictive and a functional role in determining human pluripotent stem cell neurogenic differentiation behavior. [mRNA profiling (Illumina)]: Four human ESC lines (H9, I4, I6, HUES6) at undifferentiation stages were purified with stem cell surface marker SSEA4 and subjected to RNA extraction and hybridization on Illumina microarrays. Each sample has 3 biological repeats, one of which has two technical repeats. [miRNA profiling (Agilent)]: Four human ESC lines (H9, I4, I6, HUES6) at undifferentiation stages were purified with stem cell surface marker SSEA4 and subjected to RNA extraction and hybridization on Agilent microarrays. Each sample has 3 biological repeats, one of which has two technical repeats.

2011-06-01 | E-GEOD-27834 | biostudies-arrayexpress

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data