Induction of Pluripotency in Adult Unipotent Germline Stem Cells
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE16178 | GEO | 2009/07/29
SECONDARY ACCESSION(S): PRJNA115067
REPOSITORIES: GEO
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Project description:Mouse and human stem cells with features similar to those of embryonic stem cells have been derived from testicular cells. Although pluripotent stem cells have been obtained from defined germline stem cells (GSCs) of mouse neonatal testis, only multipotent stem cells have been obtained so far from defined cells of mouse adult testis. In this study we describe a robust and reproducible protocol for obtaining germline-derived pluripotent stem (gPS) cells from adult unipotent GSCs. Pluripotency of gPS cells was confirmed by in vitro and in vivo differentiation, including germ cell contribution and transmission. As determined by clonal analyses, gPS cells indeed originate from unipotent GSCs. We propose that the conversion process requires a GSC culture microenvironment that depends on the initial number of plated GSCs and the length of culture time.
2009-07-15 | GSE11274 | GEO
Project description:Mouse and human stem cells with features similar to those of embryonic stem cells have been derived from testicular cells. Although pluripotent stem cells have been obtained from defined germline stem cells (GSCs) of mouse neonatal testis, only multipotent stem cells have been obtained so far from defined cells of mouse adult testis. In this study we describe a robust and reproducible protocol for obtaining germline-derived pluripotent stem (gPS) cells from adult unipotent GSCs. Pluripotency of gPS cells was confirmed by in vitro and in vivo differentiation, including germ cell contribution and transmission. As determined by clonal analyses gPS cells indeed originate from unipotent GSCs. We propose that the conversion process requires a GSC culture microenvironment that depends on the initial number of plated GSCs and the length of culture time. Nine samples were analyzed. GSC1: Mouse Germ Stem Cells, line 1 (1 replicate), GSC2: Mouse Germ Stem Cells, line 2 (1 replicate), GSC3: Mouse Germ Stem Cells, line 3 (1 replicate), GSC4: Mouse Germ Stem Cells, line 4 (1 replicate) gPS1: Mouse clonal germ Pluripotent Stem cells, line 1, GFP-sorted (1 replicate), gPS2: Mouse clonal germ Pluripotent Stem cells, line 2, GFP-sorted (1 replicate), gPS3: Mouse clonal germ Pluripotent Stem cells, line 3, GFP-sorted (1 replicate) ESC: Mouse Embryonic Stem Cells (duplicate)
2010-04-09 | E-GEOD-16178 | biostudies-arrayexpress
Project description:In regenerative medicine, histocompatibility of pluripotent stem cells is required to solve the problem of immunorejection after therapeutic transplantation. In this study, we show that autologous germline stem cells (GSCs), often called spermatogonial stem cells, could be derived by testis biopsy from individual mice and that GSCs subsequently could be dedifferentiated into autologous germline-derived pluripotent stem (gPS) cells. The establishment of GSCs by testicular biopsy in the mouse model can prove the principle of human clinical application to derive autologous GSCs to generate patient-specific pluripotent cells for regenerative medicine.
2011-01-23 | GSE20031 | GEO
Project description:In regenerative medicine, histocompatibility of pluripotent stem cells is required to solve the problem of immunorejection after therapeutic transplantation. In this study, we show that autologous germline stem cells (GSCs), often called spermatogonial stem cells, could be derived by testis biopsy from individual mice and that GSCs subsequently could be dedifferentiated into autologous germline-derived pluripotent stem (gPS) cells. The establishment of GSCs by testicular biopsy in the mouse model can prove the principle of human clinical application to derive autologous GSCs to generate patient-specific pluripotent cells for regenerative medicine. For transcriptome profiling, 400 ng of total DNA-free RNA was used as input for labelled cRNA synthesis (Illumina TotalPrep RNA Amplification Kit - Ambion) following the manufacturer's instructions (IVT: 12h). Quality-checked cRNA samples were hybridized as biological or technical duplicates for 18 h onto mouse-8 v2 expression BeadChips (Illumina), washed, stained, and scanned following guidelines and using materials / instrumentation supplied / suggested by the manufacturer 13 samples were analyzed, GSC1: Mouse Germ Stem Cells OG2, line 1 (1 replicate); GSC2: Mouse Germ Stem Cells OG2, line 2 (1 replicate); GSC3: Mouse Germ Stem Cells OG2, line 3 (1 replicate); GSCr1: Mouse Germ Stem Cells OG2-ROSA, line 1 (1 replicate); GSCr2: Mouse Germ Stem Cells OG2-ROSA, line 2 (2 replicate); GSCr3: Mouse Germ Stem Cells OG2-ROSA, line 3 (1 replicate); ESC: Mouse Embryonic Stem Cells (duplicate) gPS1: Mouse clonal germ Pluripotent Stem cells from OG2, line 1, (1 replicate); gPS2: Mouse clonal germ Pluripotent Stem cells from OG2, line 2, (1 replicate); gPS3: Mouse clonal germ Pluripotent Stem cells from OG2, line 3, (1 replicate); gPSr1: Mouse clonal germ Pluripotent Stem cells from OG2-ROSA, line 1, (1 replicate); gPSr2: Mouse clonal germ Pluripotent Stem cells from OG2-ROSA, line 2, (1 replicate); gPSr3: Mouse clonal germ Pluripotent Stem cells from OG2-ROSA, line 3, (1 replicate).
2011-01-23 | E-GEOD-20031 | biostudies-arrayexpress
Project description:Although reported more than once, the generation of pluripotent stem cell populations from human testis remains questionable. We recently showed that the so - called human adult germline stem cells (haGSCs), which were claimed to be pluripotent, showed a gene expression profile comparable to human fibroblasts, suggesting that these cells are originated from a fibroblastic cells. We could derive human testicular fibroblast cells (hTFCs) from several routine testicular biopsies, naming them human testicular fibroblastic cells (hTFCs). The true conversion of unipotent mouse GSCs, also known as spermatogonial stem cells into pluripotent stem cells was recently shown. However, this conversion was not observed for human cells so far. We suggest that hTFCs isolated from routine biopsies can help in the understanding, modeling and also regenerative medicine for various diseases after reprogramming them to human induced pluripotent stem cells (iPS). We successfully reprogrammed hTFCs into human iPS cells by transducing them retrovirally with reprogramming factors as already demonstrated for human dermal fibroblasts. These hTFCs derived iPS cells are comparable with human embryonic stem cells from global gene expression, promoter methylation, and they show pluripotency judged by in vitro and in vivo differentiation.
2011-04-02 | GSE21179 | GEO
Project description:Although reported more than once, the generation of pluripotent stem cell populations from human testis remains questionable. We recently showed that the so - called human adult germline stem cells (haGSCs), which were claimed to be pluripotent, showed a gene expression profile comparable to human fibroblasts, suggesting that these cells are originated from a fibroblastic cells. We could derive human testicular fibroblast cells (hTFCs) from several routine testicular biopsies, naming them human testicular fibroblastic cells (hTFCs). The true conversion of unipotent mouse GSCs, also known as spermatogonial stem cells into pluripotent stem cells was recently shown. However, this conversion was not observed for human cells so far. We suggest that hTFCs isolated from routine biopsies can help in the understanding, modeling and also regenerative medicine for various diseases after reprogramming them to human induced pluripotent stem cells (iPS). We successfully reprogrammed hTFCs into human iPS cells by transducing them retrovirally with reprogramming factors as already demonstrated for human dermal fibroblasts. These hTFCs derived iPS cells are comparable with human embryonic stem cells from global gene expression, promoter methylation, and they show pluripotency judged by in vitro and in vivo differentiation. For transcriptome profiling, 400 ng of total DNA-free RNA was used as input for labelled cRNA synthesis (Illumina TotalPrep RNA Amplification Kit - Ambion) following the manufacturer's instructions (IVT: 12h). Quality-checked cRNA samples were hybridized as biological or technical duplicates for 18 h onto human-8 v3 expression BeadChips (Illumina), washed, stained, and scanned following guidelines and using materials/instrumentation supplied/suggested by the manufacturer. 8 samples were analyzed: NCL: NCL3 hESCs grown on feeders, bulk harvest at semi-sub confluence (1 replicate) HuES6: HuES6 hESCs grown on feeders, bulk harvest at semi-sub confluence (1 replicate) HT3: Human fibroblast-like testis cells, line 3 (1 replicate) HT16: Human fibroblast-like testis cells, line 16 (1 replicate) FS: FS human foreskin fibroblasts (1 replicate) BJ: BJ human fibroblasts (1 replicate) iPS3: hiPSC line 2 derived from HT3 (1 replicate) iPS16: hiPSC line 2 derived from HT16 (1 replicate)
2011-04-02 | E-GEOD-21179 | biostudies-arrayexpress
Project description:Somatic cells can be reprogrammed to pluripotency using different methods. In comparison to pluripotent cells obtained through somatic nuclear transfer, induced pluripotent stem cells (iPSCs) exhibit a higher number of epigenetic errors. Furthermore, most of these abnormalities have been described to be intrinsic to the iPSC technology. Here we investigate whether the aberrant epigenetic patterns detected in iPSCs are specific to transcription factor-mediated reprogramming. We used germline stem cells (GSCs), which are the only adult cell type that can be converted into pluripotent cells (gPSCs) under specific culture conditions, and compared GSC-derived iPSCs and gPSCs at the transcriptomic, epigenetic and functional level. Our results show that both reprogramming methods generate indistinguishable states of pluripotency. GSC-derived iPSCs and gPSCs retained similar levels of donor cell-type memory and exhibited comparable numbers of reprogramming errors. Therefore, our study demonstrates that the epigenetic memory detected in iPSCs is not intrinsic to transcription-factor mediated reprogramming. Total RNA from 12 different in vitro mouse cell lines, 2 technical replicates per sample: germline stem cells (GSCs, 2 independent cell lines), GSC-derived induced pluripotent stem cells (iPSCs, 4 independent cell lines), germline-derived pluripotent stem cells (gPSCs, 4 independent cell lines), embryonic stem cells (ESCs), fibroblast-derived induced pluripotent stem cells (Fib-iPSCs)
2015-03-07 | E-GEOD-66613 | biostudies-arrayexpress
Project description:mouse embryonic fibroblasts, embryonic stem cells, and induced pluripotent stem cells were compared via metabolomic analysis
2018-08-02 | MSV000082680 | GNPS
Project description:The maintenance and differentiation of highly potent animal stem cells generates an epigenetic cycle that underlies development. Drosophila female germline stem cells (GSC) produce cystoblast daughters that differentiate into nurse cells and oocytes. Developmental chromatin analysis profiling the differentiation of GSCs into cystoblasts and NCs of increasing ploidy shows that cystoblasts start developing by forming heterochromatin while in a transient syncytial state, the germline cyst, reminiscent of early embryonic cells. The open GSC chromatin state is further restricted by Polycomb repression of targets that include testis expressed genes briefly active in early female germ cells. Like other highly potent stem cells, GSC metabolism is reprogrammed and Myc-dependent growth is upregulated by altering mitochondrial membrane transport, gluconeogenesis and other processes. Thus, the animal generational cycle comprises similar but distinct maternal and zygotic stem cell epigenetic cycles. We propose that the pluripotent stem cell state and daughter cell differentiation were shaped by the pressure to resist transposon activity over evolutionary time scales. In this GEO submission, we present data and analyses pertaining to H3K27ac, H3K27me3, and H3K9me3 ChIPseq, ATACseq, and RNAseq of Germline Stem Cells (GSCs) and Nurse Cells (NCs) from Drosophila melanogaster ovaries.
2023-08-16 | GSE229943 | GEO
Project description:Human primordial germ cells and mouse neonatal and adult germline stem cells are pluripotent and derive embryonic stem cell properties. Here we report the successful establishment of stable pluripotent human adult germline stem cells (haGSCs) derived from spermatogonial cells of adult human testis. Cellular and molecular characterization of haGSCs revealed many similarities to human embryonic stem (hES) cells and haGSCs produced teratomas after subcutaneous transplantation into immunodeficient mice. The haGSCs differentiated into various types of somatic cells of all three germ layers when grown under conditions used to induce the differentiation of hES cells. We conclude that the generation of haGSCs from testicular biopsies may provide simple and non-controversial access to individual cell-based therapy without the ethical and immunological problems associated with hES cells. Keywords: pluripotent stem cells characterisation
2008-05-09 | GSE11350 | GEO