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Tolerant and rejecting T cell microarray

ABSTRACT: anti-CD4, CD8 and CD40L treated versus control murine CD4+ T cells from micegrafted with hESC derived xenografts. Human embryonic stem cells (ESCs), by virtue of their capability to self-renew and differentiate into almost all body cell types, represent the first type of pluripotent stem cells (PSCs) to be used in clinical transplantation to humans in recent phase-I trials. As it is unlikely that any such cells or their progeny will survive in an allogeneic host, there is an urgent need to understand how to get long-term acceptance and functional differentiation with minimal immunosuppression. Here, we show that brief induction with combined monoclonal antibody-mediated coreceptor and costimulation blockade enables long-term survival and engraftment of murine ESCs, human ESCs, human induced PSCs, and human ESC-derived progenitor cells from all three embryonic germ layers. Tolerance induced to PSC-derived progenitors appears, remarkably, to extend to protection of their differentiated progenies, and sometimes even to different tissues derived from the same donor, suggesting linked-suppression as a likely operative mechanism. These results suggest that once tolerance has been established to PSC-derived progenitor cells, it can be extended to their differentiated progenies. Global gene expression profiling of graft infiltrating T-lymphocytes identifies gene sets that differ between rejecting and tolerant populations including gene upregulation in pathways associated with apoptosis, cell adhesion, transcription suppression and TGF synthesis; and gene downregulation in pathways associated with inflammation in recipients treated with combined coreceptor and costimulation blockade.

ORGANISM(S): Mus musculus

PROVIDER: GSE61867 | GEO | 2014/09/30

SECONDARY ACCESSION(S): PRJNA262573

REPOSITORIES: GEO

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Tolerant and rejecting T cell microarray

Project description:anti-CD4, CD8 and CD40L treated versus control murine CD4+ T cells from micegrafted with hESC derived xenografts. Human embryonic stem cells (ESCs), by virtue of their capability to self-renew and differentiate into almost all body cell types, represent the first type of pluripotent stem cells (PSCs) to be used in clinical transplantation to humans in recent phase-I trials. As it is unlikely that any such cells or their progeny will survive in an allogeneic host, there is an urgent need to understand how to get long-term acceptance and functional differentiation with minimal immunosuppression. Here, we show that brief induction with combined monoclonal antibody-mediated coreceptor and costimulation blockade enables long-term survival and engraftment of murine ESCs, human ESCs, human induced PSCs, and human ESC-derived progenitor cells from all three embryonic germ layers. Tolerance induced to PSC-derived progenitors appears, remarkably, to extend to protection of their differentiated progenies, and sometimes even to different tissues derived from the same donor, suggesting linked-suppression as a likely operative mechanism. These results suggest that once tolerance has been established to PSC-derived progenitor cells, it can be extended to their differentiated progenies. Global gene expression profiling of graft infiltrating T-lymphocytes identifies gene sets that differ between rejecting and tolerant populations including gene upregulation in pathways associated with apoptosis, cell adhesion, transcription suppression and TGF synthesis; and gene downregulation in pathways associated with inflammation in recipients treated with combined coreceptor and costimulation blockade. Mice grafted with hESC derived tissues and untreated (control) or tolerised via injections of anti-CD4, anti CD8 and CD40L. CD3+ T cells isolated from the gratfs 30 days later.

2014-09-30 | E-GEOD-61867 | biostudies-arrayexpress

Parthenogenetic stem cells for tissue-engineered heart repair

Project description:Uniparental parthenotes are considered an unwanted byproduct of in vitro fertilization. In utero parthenote development is severely compromised by defective organogenesis and in particular by defective cardiogenesis. Although developmentally compromised, apparently pluripotent stem cells can be derived from parthenogenetic blastocysts. Here we hypothesized that nonembryonic parthenogenetic stem cells (PSCs) can be directed toward the cardiac lineage and applied to tissue-engineered heart repair. We first confirmed similar fundamental properties in murine PSCs and embryonic stem cells (ESCs), despite notable differences in genetic (allelic variability) and epigenetic (differential imprinting) characteristics. Haploidentity of major histocompatibility complexes (MHCs) in PSCs is particularly attractive for allogeneic cell-based therapies. Accordingly, we confirmed acceptance of PSCs in MHC-matched allotransplantation. Cardiomyocyte derivation from PSCs and ESCs was equally effective. The use of cardiomyocyte-restricted GFP enabled cell sorting and documentation of advanced structural and functional maturation in vitro and in vivo. This included seamless electrical integration of PSC-derived cardiomyocytes into recipient myocardium. Finally, we enriched cardiomyocytes to facilitate engineering of force-generating myocardium and demonstrated the utility of this technique in enhancing regional myocardial function after myocardial infarction. Collectively, our data demonstrate pluripotency, with unrestricted cardiogenicity in PSCs, and introduce this unique cell type as an attractive source for tissue-engineered heart repair. 8 samples in total. Parthenogenetic stem cells (PSC): - PSC_1 - PSC_2 - PSC_3 Embryoid body (EB) assays of parthenogenetic stem cells: - EB_PSC_1 - EB_PSC_2 Embryonic stem cells (ESC): - ESC_1 - ESC_2 - ESC_3

2013-02-23 | E-GEOD-30868 | biostudies-arrayexpress

Generation of functional rat spermatozoa in sterile mice utilizing blastocyst complementation with pluripotent stem cells [scRNAseq_mouse]

Project description:Pluripotent stem cells (PSCs) provide a powerful tool to produce transgenic animals for biomedical research. However, impaired PSC contribution to chimerism and most notably the germline oftentimes impedes production of genetically modified animals, rendering techniques that expediate PSC contribution to the germline highly desirable. Blastocyst complementation denotes a technique which purposes to generate organs, tissues or cells in animal chimeras via microinjection of PSCs into genetically compromised blastocyst-stage embryos. Here we report on successful blastocyst complementation of the male germline in adult chimeras following microinjection of mouse or rat PSCs into mouse blastocysts mutated for Tsc22d3, an essential gene for spermatozoa production. Microinjection of mouse embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs) into Tsc22d3-KnockOut (KO) blastocyst-stage embryos gave rise to intraspecies chimeras embodying functional spermatozoa, which were solely derived from microinjected PSCs. Furthermore, microinjection of rat ESCs into Tsc22d3-KO mouse embryos gave rise to viable mouse-rat chimeras that exhibited extensive contribution of rat cells to various organs. Notably, multiple mouse-rat chimeras showed contribution of rat ESCs to the male germline, solely harboring rat spermatids and spermatozoa that were rat ESC-derived and could fertilize rat oocytes. Collectively, this study reports a method for exclusive production of functional germ cells of one species in another via blastocyst complementation with PSCs. Implications of this study extend to development of transgenic rats via sterile mice, and may further assist efforts to generate xenogeneic gametes from endangered animal species.

2022-08-01 | GSE167434 | GEO

Generation of functional rat spermatozoa in sterile mice utilizing blastocyst complementation with pluripotent stem cells [bulk RNA-seq]

2022-08-01 | GSE167433 | GEO

Generation of functional rat spermatozoa in sterile mice utilizing blastocyst complementation with pluripotent stem cells [scRNAseq_chimera]

2022-08-01 | GSE202699 | GEO

Generation of functional rat spermatozoa in sterile mice utilizing blastocyst complementation with pluripotent stem cells [scRNAseq_rat]

Project description:Pluripotent stem cells (PSCs) provide a powerful tool to produce transgenic animals for biomedical research. However, impaired PSC contribution to chimerism and most notably to the germline oftentimes impedes production of genetically modified animals, rendering techniques that expediate PSC contribution to the germline highly desirable. Blastocyst complementation denotes a technique which purposes to generate organs, tissues or cells in animal chimeras following microinjection of PSCs into genetically compromised blastocyst-stage embryos. Here, we report on successful blastocyst complementation of the male germline in adult chimeras via microinjection of mouse or rat PSCs into mouse blastocysts mutated for Tsc22d3, an essential gene for spermatozoa production. Microinjection of mouse embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs) into Tsc22d3-KnockOut (KO) blastocyst-stage embryos gave rise to intraspecies chimeras embodying functional spermatozoa, which were solely derived from the microinjected PSCs. Furthermore, microinjection of rat ESCs into Tsc22d3-KO mouse embryos gave rise to viable mouse-rat chimeras that exhibited extensive contribution of rat cells to various tissues and organs. Notably, multiple mouse-rat chimeras showed contribution of rat ESCs to the male germline, solely harboring rat spermatids and spermatozoa that were rat ESC-derived and could fertilize rat oocytes. Collectively, this study reports on a method for exclusive germ cell production of one species in another via blastocyst complementation with PSCs. Implications of this study may extend to development of transgenic rat gametes in sterile mice and could further assist efforts to generate germ cells from endangered animal species.

2022-08-01 | GSE203350 | GEO

Primed and naïve marmoset pluripotent stem cells as a model for primate development

Project description:We derived primed pluripotent stem cell cultures (PSCs) from marmoset blastocyst using conventional human PSC culture conditions (KSR/bFGF on feeders). Naïve marmoset PSCs were established by chemical resetting. In addition, we generated forebrain-derived cells from the frontal lobe of a marmoset neonate. Individual cells were manually isolated with a mouth pipette and subjected to single-cell full-lengths transcriptome profiling using a modified version of Smart-Seq2.

2022-02-14 | E-MTAB-9349 | biostudies-arrayexpress

Parthenogenetic stem cells for tissue-engineered heart repair

2013-02-23 | GSE30868 | GEO

Defining the skeletal myogenic lineage in human pluripotent stem cell-derived teratomas

Project description:Skeletal muscle stem cells are essential to muscle homeostasis and regeneration after injury. An attractive approach to obtain these cells is via differentiation of pluripotent stem cells (PSCs). We have recently reported that teratomas derived from mouse PSCs are a rich source of skeletal muscle stem cells. Here, we showed that the teratoma formation method is also capable of producing skeletal myogenic progenitors from human PSCs. Using single-cell transcriptomics, we discovered multiple lineages in human PSC-derived teratomas. Interestingly, we observed several distinct skeletal myogenic subpopulations. Trajectory analysis revealed that these subpopulations represented progressive stages of skeletal myogenic development. We further discovered that ERBB3 and CD82 are effective surface markers for prospective isolation of the skeletal myogenic lineage in human PSC-derived teratomas. Therefore, teratoma formation provides an accessible model for obtaining human skeletal myogenic progenitors from PSCs.

2022-05-18 | GSE189985 | GEO

Bulk RNA-seq of PSCs and induced PSM cells from six different mammalian species

Project description:In this study, we performed bulk RNA-seq of pluripotent stem cells (PSCs) and induced pre-somitic mesoderm (PSM) cells of six different mammalian species. The species studied are: mouse (Mus musculus), marmoset (Callithrix jacchus), rabbit (Oryctolagus cuniculus), human (Homo sapiens), cattle (Bos taurus) and southern white rhinoceros (Ceratotherium simum). We used mouse ESCs, marmoset iPSCs, rabbit ESCs, human iPSCs, bovine ESCs and rhinoceros ESCs to induce PSM-like cells from these species following protocols already described. PSC samples were extracted under maintenance conditions. PSM samples were extracted on the day when the differentiation efficiency was higher based on the percentage of cells expressing the PSM marker TBX6. We used identical culture conditions when extracting the induced PSM cells to minimize the effect of external factors on our results. Two replicates per each cell type and species were collected for a total of 24 samples. We compared the expression levels of more than 10,000 orthologous protein-coding genes across the six species. With this, we determined that the species-specific segmentation clock periods might be derived from species-specific gene expression profiles controlling basic biological processes.

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