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PROVIDER: PRJNA1089707 | ENA |
REPOSITORIES: ENA
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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:Purpose: the goal of this study is to analyse various chicken stem cell lines and to compare them with both primary somatic fibroblasts and blastodermal cells derived from pre-gastrulating embryos Methods: NGS RNA-sequencing was performed on chicken primary embryonic fibroblasts (CEF), embryonic stem cells (cESC), on blastodermal cells of EGK-X EGK-XII stages embryos (BCs), long term in vitro cultured primordial germ cells (PGCs) as specific chicken stem cells and reprogrammed cells (1A, 1D, 3E, 3F) derived from CEF by somatic reprogramming process. The librairies were prepared using the TruSeq R Stranded mRNA sample preparation kit (Illumina). The paired-end sequencing was performed on the NextSeq 500 sequencer (Illumina) and controlled by the Seqencing Analysis viewer software (Illumina). The quality control of the Sample-ID.fastq files was performed with the FasQC software (Babraham Institute). Results: the total number of reads ranges from was of 80 millions to 145 millions.
2018-04-08 | GSE102353 | GEO
Project description:The present investigation was to identify the signaling and metabolic pathways of expressed genes by microarray comparison between Primordial Germ Cells (PGCs) and their somatic counterpart, chicken embryonic fibroblasts (CEFs). We identified a total of 2,605 expressed transcripts. Among these, 1,197 were predominantly expressed in PGCs, and 1,408 were predominantly expressed in CEFs.
2013-01-10 | GSE38168 | GEO
Project description:Chicken primordial germ cells (PGCs) have an epigenetic signature which differs from the one that mammalian PGCs acquire with their epigenome reprogramming during early embryonic development. In particular, chicken PGCs display a high global amount of histone H3 lysine 9 trimethylation (H3K9me3) compared to somatic cell types. We performed the genome-wide profiling of H3K9me3 and the transcriptome analysis on chicken PGCs compared to embryonic stem cells (ESCs) as a closely related, non germinal cell type.
2024-04-26 | GSE248767 | GEO
Project description:Chicken primordial germ cells (PGCs) have an epigenetic signature which differs from the one that mammalian PGCs acquire with their epigenome reprogramming during early embryonic development. In particular, chicken PGCs display a high global amount of histone H3 lysine 9 trimethylation (H3K9me3) compared to somatic cell types. We performed the genome-wide profiling of H3K9me3 and the transcriptome analysis on chicken PGCs compared to embryonic stem cells (ESCs) as a closely related, non germinal cell type.
2024-04-26 | GSE248766 | GEO
Project description:The comparative transcriptomic analysis of chicken stem cells defines a chicken set of pluripotency associated genes that is almost coincident with mammalian pluripotency-assocaited genes. A total of 5944 differentially expressed unique identifiers (target IDs) were defined for CEF, 5942 for BM2 cells, 4550 for cES, 5465 for PGC and 5522 for cBC. Chicken Blastodermal cells (cBC) were taken from stage IX-XII (Eyal-Giladi & Kochav, 1976) embryos,chicken embryonic stem (cES) cells were established, amplified on inactivated STO feeder cells in proliferative medium containing cytokines and growth factors as described (Pain et al., 1996, Lavial et al., 2007). Long term cultured primordial germ cells (PGCs) were derived from 48h embryonic blood and maintained as described (McDonald et al., 2010). The non-tumorigenic BM2 monocytic cell line was grown as described (Solari et al., 1996). Primary chicken Embryonic fibroblasts (CEF) were prepared from 11-12 day old embryos according to standard protocols (Gandrillon et al., 1987), maintained and homogenised for 3-4 passages before being used as a somatic cell control. RNAs were extracted using Trizol (Invitrogen) according to the manufacturer and microarray analysis performed in biological triplicate.
2014-11-01 | E-GEOD-61221 | biostudies-arrayexpress
Project description:Transcriptome analysis of chicken ES, blastodermal, primordial germ cells, embryonic fibroblasts and monocytic cells
| PRJNA260527 | ENA
Project description:The molecular mechanisms underlying the commitment of cells to the germ cell lineage and the subsequent formation of germ cells during mammalian development remain poorly understood due to an inability to obtain sufficient amounts of cellular materials to conduct thorough in-vitro analyses. Although primordial germ cells (PGCs) have been generated and isolated from embryonic stem cells (ESCs) in vitro, concurrent expression of several cell surface receptors and transcription factors, at various levels, confounds the identification of these cells. To date, only a few genes that mark the onset of germ cell commitment to the epiblast, including tissue nonspecific alkaline phosphatase (TNAP), Blimp1, Stella, and Fragilis, have been used with some degree of success to detect PGC formation in in-vitro model systems. In light of the current literature, the generation of an unlimited supply of germ cells and gametes from pluripotent stem cells would greatly facilitate studies into reproductive development. To accomplish this, we would need to discover novel markers that are specifically expressed in germ cells. Here we identified four genes that are specifically expressed in male and female germ cells, both in vivo and in vitro, but are not expressed in somatic tissues or ESCs. Expression of these genes therefore allows us to distinguish committed germ cells from undifferentiated pluripotent cell populations, a prerequisite for the successful derivation of germ cells and gametes in vitro. For RNA isolation, PGCs were sorted by FACS, collected by centrifugation, lysed in RLT buffer (QIAGEN), and processed using RNeasy Micro kits with on-column DNase digestion as per the manufacturer's instructions. Integrity of RNA samples was quality-checked using a 2100 Bioanalyzer (Agilent). If possible, 300 ng of total RNA per sample was used as starting material for linear amplification (Illumina TotalPrep RNA amplification kit, Ambion), which involved synthesis of T7-linked double-stranded cDNA and 14 hrs of in-vitro transcription incorporating biotin-labeled nucleotides. Purified and labeled cRNA was then quality-checked on a 2100 Bioanalyser, and hybridized as biological or technical duplicates for 18 hrs onto MouseRef-8 v2 gene expression BeadChips (Illumina), following the manufacturer's instructions. After being washed, the chips were stained with streptavidin-Cy3 (GE Healthcare) and scanned using the iScan reader and accompanying software. 44 samples were analyzed. ESCm: OG2 male Embryonic Stem Cell, +/+ Oct4-GFP ES cells, 1 biological rep 11.5: 11.5 embryonic day PGC (Primordial Germ Cell), 2 biological reps 12.5F: 12.5 embryonic day female PGC (Primordial Germ Cell), 2 biological reps 13.5F: 13.5 embryonic day female PGC (Primordial Germ Cell), 2 biological reps 14.5F: 14.5 embryonic day female PGC (Primordial Germ Cell), 2 biological reps 15.5F: 15.5 embryonic day female PGC (Primordial Germ Cell), 2 biological reps 16.5F: 16.5 embryonic day female PGC (Primordial Germ Cell), 2 biological reps 17.5F: 17.5 embryonic day female PGC (Primordial Germ Cell), 2 biological reps 18.5F: 18.5 embryonic day female PGC (Primordial Germ Cell), 2 biological reps 12.5M: 12.5 embryonic day male PGC (Primordial Germ Cell), 2 biological reps 13.5M: 13.5 embryonic day male PGC (Primordial Germ Cell), 2 biological reps 14.5M: 14.5 embryonic day male PGC (Primordial Germ Cell), 2 biological reps 15.5M: 15.5 embryonic day male PGC (Primordial Germ Cell), 2 biological reps 16.5M: 16.5 embryonic day male PGC (Primordial Germ Cell), 2 biological reps 17.5M: 17.5 embryonic day male PGC (Primordial Germ Cell), 2 biological reps 18.5M: 18.5 embryonic day male PGC (Primordial Germ Cell), 2 biological reps 8M: 8 day after birth male PGC (Primordial Germ Cell), 2 biological reps 10M: 10 day after birth male PGC (Primordial Germ Cell), 2 biological reps 12M: 12 day after birth male PGC (Primordial Germ Cell), 2 biological reps 14M: 14 day after birth male PGC (Primordial Germ Cell), 1 biological rep 16M: 16 day after birth male PGC (Primordial Germ Cell), 2 biological reps 18M: 18 day after birth male PGC (Primordial Germ Cell), 2 biological reps 20M: 20 day after birth male PGC (Primordial Germ Cell), 2 biological reps
2010-11-16 | E-GEOD-23322 | biostudies-arrayexpress
Project description:Generation of genetically uniformed individuals from somatic cells is an effective approach for large-scale reproduction of elite varieties and a powerful tool for restoration of endangered species. However, this technique has never been realized in avian due to their oviparous reproduction pattern. In this study, we produced cloned-like chicken by allogeneic transplantation of somatic cells induced primordial germ cells (PGCs). Oct4, Sox2, Nanog and Lin28A (OSNL) factors were employed to reprogram primary chicken embryo fibroblasts (CEF) to pluripotent stem cells (iPS), in which DNA demethylation and histone acetylation were found to increase the efficiency to 13.00%. The obtained iPS presented embryonic stem cell like characters and were further induced to PGCs by BMP4/BMP8b/EGF, in which histone acetylation and glycolysis inhibition elevated the induction rate to 17.30% and 16.41%, respectively. With the optimized system, we induced Black Langshan Chicken (Gallus domestiaus, black feather) donated CEF to PGCs and transplanted them into the Recessive White Chicken (white feather) embryos. The transplanted cells migrated into the genital ridges and produced functional sperm or oocytes. The sexual matured recipients were self-crossed, with 189/509 (37.13%) cloned-like chicken produced. Microsatellite analysis confirmed their DNA inheritance from the black donor chicken. Thus, we demonstrated, for the first time, the feasibility of avian cloning from somatic cells.
2021-03-16 | GSE159511 | GEO