Similar Datasets
Project description:The ability to generate synthetic mouse embryo models (SEMs) that can grow beyond gastrulation and early organogenesis has proven the plasticity of naïve Pluripotent stem cells (nPSC) to fully recapitulate the developmental dynamics of the mouse embryo. Existing protocols rely on the induction of extraembryonic lineages by ectopically expressing master transcription factors that induce their differentiation to trophoblast (TE) or primitive endoderm (PrE) that latter will form the placenta and yolk sac, respectively. However, this approach brings technical difficulties related to the required genetic manipulation, separate lineage induction and mixing of different cell lines. While several works have suggested efficient priming to extraembryonic lineages without the need of transgenes, they have not proven the capability of this cells to contribute to the formation of SEMs. In this study, we demonstrate that naive ESCs can be co-induced in a single transgene free condition to give rise to both embryonic and extraembryonic lineages present in the preimplantation mouse embryo while preserving an epiblast compartment. Furthermore, this condition can be aggregated and cultured, resembling post-implantation egg cylinders, and by applying ex-utero growth technologies they progress to mimic features corresponding to E8.5 in-utero growth embryos. Moreover, by optimization of a media termed Alternative Condition (AC), we provide ready to use condition that can be maintained long term and directly aggregated to generate TF-SEMs, eliminating the need of cell preinductions before aggregation.
2025-07-29 | GSE294685 | GEO
Project description:The ability to generate Synthetic mouse Embryo Models (SEMs) that can grow beyond gastrulation and early organogenesis has proven the plasticity of naïve Pluripotent stem cells (nPSC) to fully recapitulate the developmental dynamics of the mouse embryo. Existing protocols rely on the induction of extraembryonic lineages by ectopically expressing master transcription factors that induce their differentiation to trophoblast (TE) or primitive endoderm (PrE) that latter will form the placenta and yolk sac, respectively. However, this approach brings technical difficulties related to the required genetic manipulation, separate lineage induction and mixing of nPSC. While several works have suggested efficient priming to extraembryonic lineages without the need of transgenes, they have not proven the capability of this cells to contribute to the formation of SEMs. In this study, we demonstrate that nPSCs can be co-induced in a single transgene free condition to give rise to both embryonic and extraembryonic lineages present in the preimplantation mouse embryo while preserving an epiblast compartment. Furthermore, this condition can be aggregated and cultured, resembling post-implantation egg cylinders, and by applying ex-utero growth technologies they progress to mimic features corresponding to E8.5 in-utero growth embryos. Moreover, by optimization of a media termed Alternative Pluripotent Condition (APC), we provide ready to use condition that can be maintained long term and directly aggregated to generate TF-SEMs, eliminating the need of cell preinductions before aggregation.
2025-07-29 | GSE294690 | GEO
Project description:Transgene-free generation of mouse post-gastrulation whole embryo models solely from naive ESCs and iPSCs
| PRJNA1251571 | ENA
Project description:In vitro cultured stem cells with distinct developmental capacities can contribute to embryonic or extraembryonic tissues after microinjection into pre-implantation mammalian embryos. However, whether cultured stem cells can independently give rise to entire gastrulating embryo-like structures with embryonic and extraembryonic compartments remains unknown. Here, we adapt a recently established platform for prolonged ex utero growth of natural embryos to generate mouse post-gastrulation synthetic whole embryo models (sEmbryos), with both embryonic and extraembryonic compartments, starting solely from naive ESCs. This was achieved by co-aggregating non-transduced ESCs, with naive ESCs transiently expressing Cdx2 or Gata4 to promote their priming toward trophectoderm and primitive endoderm lineages, respectively. sEmbryos adequately accomplish gastrulation, advance through key developmental milestones, and develop organ progenitors within complex extraembryonic compartments similar to E8.5 stage mouse embryos. Our findings highlight the plastic potential of naive pluripotent cells to self-organize and functionally reconstitute and model the entire mammalian embryo beyond gastrulation.
| S-EPMC9439721 | biostudies-literature
Project description:Signaling Induced Plasticity Enables Transgene-Free Generation of Mouse Post-Gastrulation Whole Embryo Models Solely from Naïve ESCs and iPSCs (ii)
| PRJNA1251095 | ENA
Project description:Signaling Induced Plasticity Enables Transgene-Free Generation of Mouse Post-Gastrulation Whole Embryo Models Solely from Naïve ESCs and iPSCs (i)
| PRJNA1251082 | ENA
Project description:The ability to study human post-implantation development remains limited owing to ethical and technical challenges associated with intrauterine development after implantation1. Embryo-like models with spatially organized morphogenesis and structure of all defining embryonic and extra-embryonic tissues of the post-implantation human conceptus (that is, the embryonic disc, the bilaminar disc, the yolk sac, the chorionic sac and the surrounding trophoblast layer) remain lacking1,2. Mouse naive embryonic stem cells have recently been shown to give rise to embryonic and extra-embryonic stem cells capable of self-assembling into post-gastrulation structured stem-cell-based embryo models with spatially organized morphogenesis (called SEMs)3. Here we extend those findings to humans using only genetically unmodified human naive embryonic stem cells (cultured in human enhanced naive stem cell medium conditions)4. Such human fully integrated and complete SEMs recapitulate the organization of nearly all known lineages and compartments of post-implantation human embryos, including the epiblast, the hypoblast, the extra-embryonic mesoderm and the trophoblast layer surrounding the latter compartments. These human complete SEMs demonstrated developmental growth dynamics that resemble key hallmarks of post-implantation stage embryogenesis up to 13-14 days after fertilization (Carnegie stage 6a). These include embryonic disc and bilaminar disc formation, epiblast lumenogenesis, polarized amniogenesis, anterior-posterior symmetry breaking, primordial germ-cell specification, polarized yolk sac with visceral and parietal endoderm formation, extra-embryonic mesoderm expansion that defines a chorionic cavity and a connecting stalk, and a trophoblast-surrounding compartment demonstrating syncytium and lacunae formation. This SEM platform will probably enable the experimental investigation of previously inaccessible windows of human early post implantation up to peri-gastrulation development.
| S-EPMC10584686 | biostudies-literature
Project description:Non-human primates (NHPs) are pivotal animal models for translating novel cell replacement therapies into clinical applications, including validating the safety and efficacy of induced pluripotent stem cell (iPSC)-derived products. Preclinical development and the testing of cell-based therapies ideally comprise xenogeneic (human stem cells into NHPs) and allogenic (NHP stem cells into NHPs) transplantation studies. For the allogeneic approach, it is necessary to generate NHP-iPSCs with generally equivalent quality to the human counterparts that will be used later on in patients. Here, we report the generation and characterization of transgene- and feeder-free cynomolgus monkey (Macaca fascicularis) iPSCs (Cyno-iPSCs). These novel cell lines have been generated according to a previously developed protocol for the generation of rhesus macaque, baboon, and human iPSC lines. Beyond their generation, we demonstrate the potential of the novel Cyno-iPSCs to differentiate into two clinically relevant cell types, i.e., cardiomyocytes and neurons. Overall, we provide a resource of novel iPSCs from the most frequently used NHP species in the regulatory testing of biologics and classical pharmaceutics to expand our panel of iPSC lines from NHP species with high relevance in preclinical testing and translational research.
| S-EPMC10969578 | biostudies-literature
Project description:A variety of somatic cells can be reprogrammed to induced pluripotent stem cells (iPSCs), but CD34(+) hematopoietic stem cells (HSCs) present in nonmobilized peripheral blood (PB) would be a convenient target. We report a method for deriving iPSC from PB HSCs using immunobead purification and 2- to 4-day culture to enrich CD34(+) HSCs to 80% ± 9%, followed by reprogramming with loxP-flanked polycistronic (human Oct4, Klf4, Sox2, and c-Myc) STEMCCA-loxP lentivector, or with Sendai vectors. Colonies arising with STEMCCA-loxP were invariably TRA-1-60(+), yielding 5.3 ± 2.8 iPSC colonies per 20 mL PB (n = 17), where most colonies had single-copy STEMCCA-loxP easily excised by transient Cre expression. Colonies arising with Sendai were variably reprogrammed (10%-80% TRA-1-60(+)), with variable yield (6 to >500 TRA-1-60(+) iPSC colonies per 10 mL blood; n = 6). Resultant iPSC clones expressed pluripotent cell markers and generated teratomas. Genomic methylation patterns of STEMCCA-loxP-reprogrammed clones closely matched embryonic stem cells. Furthermore, we showed that iPSCs are derived from the nonmobilized CD34(+) HSCs enriched from PB rather than from any lymphocyte or monocyte contaminants because they lack somatic rearrangements typical of T or B lymphocytes and because purified CD14(+) monocytes do not yield iPSC colonies under these reprogramming conditions.
| S-EPMC3617640 | biostudies-literature
Project description:Induced pluripotent stem cells (iPSCs) are capable of providing an unlimited source of cells from all three germ layers and germ cells. The derivation and usage of iPSCs from various animal models may facilitate stem cell-based therapy, gene-modified animal production, and evolutionary studies assessing interspecies differences. However, there is a lack of species-wide methods for deriving iPSCs, in particular by means of non-viral and non-transgene-integrating (NTI) approaches. Here, we demonstrate the iPSC derivation from somatic fibroblasts of multiple mammalian species from three different taxonomic orders, including the common marmoset (Callithrix jacchus) in Primates, the dog (Canis lupus familiaris) in Carnivora, and the pig (Sus scrofa) in Cetartiodactyla, by combinatorial usage of chemical compounds and NTI episomal vectors. Interestingly, the fibroblasts temporarily acquired a neural stem cell-like state during the reprogramming. Collectively, our method, robustly applicable to various species, holds a great potential for facilitating stem cell-based research using various animals in Mammalia.
| S-EPMC8072067 | biostudies-literature