Project description:The molecular characteristics of EPSCs have long been debated. While human EPSCs (hEPSCs) resemble eight-cell and morula-stage embryos, mouse EPSCs (mEPSCs) more closely resemble post-implantation epiblasts. This study explores the developmental potential and molecular characteristics of porcine expanded pluripotent stem cells (pEPSCs), uncovering their unique transcriptional heterogeneity and similarities to both morula and post-implantation epiblast stages. By performing single-cell transcriptomic analysis, three distinct subpopulations (C1, C2, and C3) were identified, corresponding to totipotency, naïve pluripotency, and primed pluripotency. The use of dual fluorescence reporter systems highlighted transitions between these states, resembling embryonic development. Key findings reveal a dynamic balance model of LEUTX and OTX2 in regulating pluripotency transition that OTX2 promotes the shift from totipotency to primed pluripotency by promoting pluripotency and lineage-priming genes, while LEUTX enhances totipotency by activating totipotency genes. LEUTX-overexpressing cells demonstrated enhanced developmental potential and stability in vitro. The study established a conserved framework for understanding state transitions mediated by OTX2 and LEUTX, with implications for regenerative medicine, developmental biology, and highlight the potential of pEPSCs in agricultural and biomedical applications.
Project description:Extended pluripotent or expanded potential stem cells (EPSCs) possess the superior developmental potential to embryonic stem cells (ESCs). However, the molecular underpinning of their in vitro maintenance is not well defined. We comparatively studied transcriptome, chromatin accessibility, active histone modification chromatin marks and relative proteomes of ESCs and the two well-established EPSCs to probe the molecular foundation of their unique developmental potential. We found a great overlap in the transcriptomic and chromatin accessibility profiles and reliance on pluripotency factors Oct4, Sox2, and Nanog for self-renewal between ESCs and EPSCs. Importantly, we identified a subset of genomic, transcriptomic, and proteomic signatures that distinguish EPSCs from ESCs in transcriptional and translational regulation as well as epigenetic and metabolic control. We also identified molecular differences between the two well-established EPSCs. Our study provides a rich resource for dissecting the regulatory network underlying the developmental potency of EPSCs and exploring alternative states of totipotency.
Project description:Extended pluripotent or expanded potential stem cells (EPSCs) possess the superior developmental potential to embryonic stem cells (ESCs). However, the molecular underpinning of their in vitro maintenance is not well defined. We comparatively studied transcriptome, chromatin accessibility, active histone modification chromatin marks and relative proteomes of ESCs and the two well-established EPSCs to probe the molecular foundation of their unique developmental potential. We found a great overlap in the transcriptomic and chromatin accessibility profiles and reliance on pluripotency factors Oct4, Sox2, and Nanog for self-renewal between ESCs and EPSCs. Importantly, we identified a subset of genomic, transcriptomic, and proteomic signatures that distinguish EPSCs from ESCs in transcriptional and translational regulation as well as epigenetic and metabolic control. We also identified molecular differences between the two well-established EPSCs. Our study provides a rich resource for dissecting the regulatory network underlying the developmental potency of EPSCs and exploring alternative states of totipotency.
Project description:Extended pluripotent or expanded potential stem cells (EPSCs) possess the superior developmental potential to embryonic stem cells (ESCs). However, the molecular underpinning of their in vitro maintenance is not well defined. We comparatively studied transcriptome, chromatin accessibility, active histone modification chromatin marks and relative proteomes of ESCs and the two well-established EPSCs to probe the molecular foundation of their unique developmental potential. We found a great overlap in the transcriptomic and chromatin accessibility profiles and reliance on pluripotency factors Oct4, Sox2, and Nanog for self-renewal between ESCs and EPSCs. Importantly, we identified a subset of genomic, transcriptomic, and proteomic signatures that distinguish EPSCs from ESCs in transcriptional and translational regulation as well as epigenetic and metabolic control. We also identified molecular differences between the two well-established EPSCs. Our study provides a rich resource for dissecting the regulatory network underlying the developmental potency of EPSCs and exploring alternative states of totipotency. This SuperSeries is composed of the SubSeries listed below.
Project description:Despite intensive efforts, establishing porcine embryonic stem cells have been challenging. We recently derived mouse expanded potential stem cells (EPSCs) from individual blastomeres by inhibiting the activity of critical molecular pathways that predisposes lineage differentiation in the mouse preimplantation embryo. EPSCs had enriched molecular signatures of blastomeres and possessed the developmental potency to all embryonic and extraembryonic cell lineages. In this study, we report the derivation of porcine EPSC (pEPSC) lines either directly from preimplantation embryos or by reprogramming fetal fibroblasts. Under similar culture conditions, human ESCs and iPSCs can be converted, or somatic cells are directly reprogrammed, to EPSCs (hEPSCs) that display the molecular and functional attributes reminiscent of pEPSCs. Here, we performed Single-Cell RNA-seq experiments to characterise the transcriptional heterogeneity of the EPSCs.
Project description:Extended pluripotent or expanded potential stem cells (EPSCs) possess the superior developmental potential to embryonic stem cells (ESCs). However, the molecular underpinning of their in vitro maintenance is not well defined. We comparatively studied transcriptome, chromatin accessibility, active histone modification chromatin marks and relative proteomes of ESCs and the two well-established EPSCs to probe the molecular foundation of their unique developmental potential. We found a great overlap in the transcriptomic and chromatin accessibility profiles and reliance on pluripotency factors Oct4, Sox2, and Nanog for self-renewal between ESCs and EPSCs. Importantly, we identified a subset of genomic, transcriptomic, and proteomic signatures that distinguish EPSCs from ESCs in transcriptional and translational regulation as well as epigenetic and metabolic control. We also identified molecular differences between the two well-established EPSCs. Our study provides a rich resource for dissecting the regulatory network underlying the developmental potency of EPSCs and exploring alternative states of totipotency.
Project description:Despite intensive efforts, establishing porcine embryonic stem cells have been challenging. We recently derived mouse expanded potential stem cells (EPSCs) from individual blastomeres by inhibiting the activity of critical molecular pathways that predisposes lineage differentiation in the mouse preimplantation embryo. EPSCs had enriched molecular signatures of blastomeres and possessed the developmental potency to all embryonic and extraembryonic cell lineages. In this study, we report the derivation of porcine EPSC (pEPSC) lines either directly from preimplantation embryos or by reprogramming fetal fibroblasts. Under similar culture conditions, human ESCs and iPSCs can be converted, or somatic cells are directly reprogrammed, to EPSCs (hEPSCs) that display the molecular and functional attributes reminiscent of pEPSCs. Here, we performed ChIP-seq experiments of H3K4me3 and H3K27me3 to characterise the epigenetic signatures of the EPSCs.
Project description:Despite intensive efforts, establishing porcine embryonic stem cells have been challenging. We recently derived mouse expanded potential stem cells (EPSCs) from individual blastomeres by inhibiting the activity of critical molecular pathways that predisposes lineage differentiation in the mouse preimplantation embryo. EPSCs had enriched molecular signatures of blastomeres and possessed the developmental potency to all embryonic and extraembryonic cell lineages. In this study, we report the derivation of porcine EPSC (pEPSC) lines either directly from preimplantation embryos or by reprogramming fetal fibroblasts. Under similar culture conditions, human ESCs and iPSCs can be converted, or somatic cells are directly reprogrammed, to EPSCs (hEPSCs) that display the molecular and functional attributes reminiscent of pEPSCs. Here, we performed RNA-seq experiments to characterise the transcriptional signatures of the EPSCs.
Project description:Totipotent embryos lack higher-order genome architecture. In this study, we profile the 3D genome architecture of two putative totipotent cellular models, ciTotiSCs and TBLCs. Notably, both ciTotiSCs and TBLCs retain TADs and structurally resemble ICM/ESCs more closely than totipotent embryos. While TADs and compartmentalization are positionally conserved, they weaken upon acquisition of totipotency. Integrative analysis of epigenetic and Hi-C data further reveal that pluripotency genes undergo coordinated epigenetic landscape remodeling and contact domain reorganization, driving pluripotency suppression. Conversely, totipotency gene activation appears to operate through distinct, yet undefined mechanisms. This study establishes the first functional link between 3D genome folding and totipotency-pluripotency transitions while proposing a revised framework for evaluating in vitro totipotency models.
Project description:Totipotent embryos lack higher-order genome architecture. In this study, we profile the 3D genome architecture of two putative totipotent cellular models, ciTotiSCs and TBLCs. Notably, both ciTotiSCs and TBLCs retain TADs and structurally resemble ICM/ESCs more closely than totipotent embryos. While TADs and compartmentalization are positionally conserved, they weaken upon acquisition of totipotency. Integrative analysis of epigenetic and Hi-C data further reveal that pluripotency genes undergo coordinated epigenetic landscape remodeling and contact domain reorganization, driving pluripotency suppression. Conversely, totipotency gene activation appears to operate through distinct, yet undefined mechanisms. This study establishes the first functional link between 3D genome folding and totipotency-pluripotency transitions while proposing a revised framework for evaluating in vitro totipotency models.