Capture of germ-line competent stem cells between rosette-stage and formative pluripotent states
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ABSTRACT:
PROVIDER: PRJNA1121218 | ENA |
REPOSITORIES: ENA
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Project description:The establishment of stem cells with varying pluripotency potentials is crucial for understanding the developmental progression of pre- and post-implantation embryos. Despite the advancement through the proposal of the formative pluripotent state, the model of pluripotent development remains incomplete. Here, we derived a novel type of pluripotent stem cells from mouse embryonic stem cells and sustained them in vitro through the application of MEK inhibitor PD0325901, Tankyrase inhibitor IWR1, and adenylate cyclase activator Forskolin. These cells exhibit distinct transcriptome and epigenetic profiles, resembling the epiblasts of E5.0-E5.5, and reside between rosette-like stem cells and formative stem cells. We refer to them as rfISCs (Intermediate stem cells between RSCs and FSCs). rfISCs can differentiate into germ cells in vitro and form blastocyst chimeras in vivo. Additionally, the formation of rfISCs is governed by distinct signaling pathways and transcription factors. Specifically, IWR1-mediated inhibition of Wnt signaling enhances Tcf7l1-mediated Otx2 expression, essential for rfISCs generation yet prone to inducing neural differentiation, whereas Forskolin induces Id1 transcription, suppressing neural lineage gene expression via the PKA pathway. Hence, co-overexpression of Id1 and Otx2 is sufficient to sustain rfISCs stability. The emergence of rfISCs aids in comprehending early embryo development.
2024-06-30 | GSE269324 | GEO
Project description:The establishment of stem cells with varying pluripotency potentials is crucial for understanding the developmental progression of pre- and post-implantation embryos. Despite the advancement through the proposal of the formative pluripotent state, the model of pluripotent development remains incomplete. Here, we derived a novel type of pluripotent stem cells from mouse embryonic stem cells and sustained them in vitro through the application of MEK inhibitor PD0325901, Tankyrase inhibitor IWR1, and adenylate cyclase activator Forskolin. These cells exhibit distinct transcriptome and epigenetic profiles, resembling the epiblasts of E5.0-E5.5, and reside between rosette-like stem cells and formative stem cells. We refer to them as rfISCs (Intermediate stem cells between RSCs and FSCs). rfISCs can differentiate into germ cells in vitro and form blastocyst chimeras in vivo. Additionally, the formation of rfISCs is governed by distinct signaling pathways and transcription factors. Specifically, IWR1-mediated inhibition of Wnt signaling enhances Tcf7l1-mediated Otx2 expression, essential for rfISCs generation yet prone to inducing neural differentiation, whereas Forskolin induces Id1 transcription, suppressing neural lineage gene expression via the PKA pathway. Hence, co-overexpression of Id1 and Otx2 is sufficient to sustain rfISCs stability. The emergence of rfISCs aids in comprehending early embryo development.
2024-07-30 | GSE270044 | GEO
Project description:Capture of germ-line competent stem cells between rosette-stage and formative pluripotent states [CUT&Tag]
| PRJNA1124906 | ENA
Project description:Naive pluripotent cells in the implanting mouse blastocyst generate a rosette structure before undergoing lumenogenesis to form the egg cylinder. Simultaneously, they acquire primed pluripotency, the ability to differentiate into the primary germ layers. The existence of discrete intermediate pluripotent states during this transition has not been demonstrated. We identify here a distinct rosette pluripotent state, defined by co-expression of naive factors with transcription factor OTX2. Downregulation of WNT signals in the blastocyst drives transition into rosette pluripotency by inducing OTX2. The rosette then activates MEK signals that induce lumenogenesis and drive progression to primed pluripotency. Consequently, combined WNT and MEK inhibition supports rosette-like stem cells (RSCs), a self-renewing naive-primed intermediate. RSCs gain a unique epigenome that includes erasure of constitutive heterochromatin and bivalent marking of primed pluripotency genes. Notwithstanding this primed chromatin landscape, WNT induces reversion to naive pluripotency. The rosette is therefore a reversible pluripotent intermediate where control over pluripotency progression and morphogenesis pivots from WNT to MEK signals.
2020-04-28 | GSE105762 | GEO
Project description:Naive pluripotent cells in the implanting mouse blastocyst generate a rosette structure before undergoing lumenogenesis to form the egg cylinder. Simultaneously, they acquire primed pluripotency, the ability to differentiate into the primary germ layers. The existence of discrete intermediate pluripotent states during this transition has not been demonstrated. We identify here a distinct rosette pluripotent state, defined by co-expression of naive factors with transcription factor OTX2. Downregulation of WNT signals in the blastocyst drives transition into rosette pluripotency by inducing OTX2. The rosette then activates MEK signals that induce lumenogenesis and drive progression to primed pluripotency. Consequently, combined WNT and MEK inhibition supports rosette-like stem cells (RSCs), a self-renewing naive-primed intermediate. RSCs gain a unique epigenome that includes erasure of constitutive heterochromatin and bivalent marking of primed pluripotency genes. Notwithstanding this primed chromatin landscape, WNT induces reversion to naive pluripotency. The rosette is therefore a reversible pluripotent intermediate where control over pluripotency progression and morphogenesis pivots from WNT to MEK signals.
2020-04-28 | GSE145726 | GEO
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 | GSE135989 | GEO
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
Project description:Primordial germ cell like cell competent populations
| PRJEB48665 | ENA
Project description:Upon implantation, the naive pluripotent epiblast of the mouse blastocyst generates a rosette, undergoes lumenogenesis and forms the primed pluripotent egg cylinder, able to generate the embryonic tissues. How pluripotency progression and morphogenesis are linked, and whether intermediate pluripotent states exist remain controversial. We identify here a rosette pluripotent state, defined by co-expression of naive factors with transcription factor OTX2. Downregulation of blastocyst WNT signals drives transition into rosette pluripotency by inducing OTX2. The rosette then activates MEK signals that induce lumenogenesis and drive progression to primed pluripotency. Consequently, combined WNT and MEK inhibition supports rosette-like stem cells (RSCs), a self-renewing naive-primed intermediate. RSCs erase constitutive heterochromatin marks and display a primed chromatin landscape, with bivalently marked primed pluripotency genes. Nonetheless, WNT induces reversion to naive pluripotency. The rosette is therefore a reversible pluripotent intermediate where control over both pluripotency progression and morphogenesis pivots from WNT to MEK signals.
2020-04-28 | GSE112234 | GEO
Project description:Pluripotent cells emerge as a naïve founder population in the blastocyst, acquire capacity for germline and soma formation, and then undergo lineage priming. Mouse embryonic stem (ES) cells and epiblast stem cells (EpiSCs) respectively represent the initial naïve and final primed phases of pluripotency. Here we investigated the intermediate formative stage. Using minimal exposure to specification cues, we derived stem cells from formative mouse epiblast. Unlike ES cells or EpiSCs, formative stem (FS) cells responded directly to germ cell induction. They colonised somatic tissues and germline in chimaeras. Whole transcriptome analyses showed similarity to pre-gastrulation formative epiblast. Signal responsiveness and chromatin accessibility features reflect lineage capacitation. Furthermore, FS cells showed distinct transcription factor dependencies, relying critically on Otx2. Finally, FS cell culture conditions applied to human naïve cells or embryos supported expansion of similar stem cells, consistent with a conserved staging post on the trajectory of mammalian pluripotency. This SuperSeries is composed of the SubSeries listed below.
2020-12-04 | GSE156589 | GEO