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NANOG is required for the long-term establishment of avian somatic reprogrammed cells

ABSTRACT: This SuperSeries is composed of the SubSeries listed below.

ORGANISM(S): Gallus gallus Anas platyrhynchos

PROVIDER: GSE109970 | GEO | 2018/04/08

REPOSITORIES: GEO

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NANOG Is Required for the Long-Term Establishment of Avian Somatic Reprogrammed Cells.

Project description:Somatic reprogramming, which was first identified in rodents, remains poorly described in non-mammalian species. Here, we generated avian reprogrammed cells by reprogramming of chicken and duck primary embryonic fibroblasts. The efficient generation of long-term proliferating cells depends on the method of delivery of reprogramming factors and the addition of NANOG and LIN28 to the canonical OCT4, SOX2, KLF4, and c-MYC gene combination. The reprogrammed cells were positive for several key pluripotency-associated markers including alkaline phosphatase activity, telomerase activity, SSEA1 expression, and specific cell cycle and epigenetic markers. Upregulated endogenous pluripotency-associated genes included POU5F3 (POUV) and KLF4, whereas cells failed to upregulate NANOG and LIN28A. However, cells showed a tumorigenic propensity when injected into recipient embryos. In conclusion, although the somatic reprogramming process is active in avian primary cells, it needs to be optimized to obtain fully reprogrammed cells with similar properties to those of chicken embryonic stem cells.

| S-EPMC6235669 | biostudies-literature

NANOG is required for the long-term establishment of avian somatic reprogrammed cells [duck]

Project description:Purpose: the goal of this study is to analyse various duck 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 duck primary embryonic fibroblasts (DEF), on blastodermal cells from EGK-X to EG XV embryos (BCs) and reprogrammed cells (R71-2 and R71-5) derived from DEF by somatic reprogramming process. The libraries were prepared using the TruSeq R Stranded mRNA sample preparation kit (Illumina). The paired-end sequencing was performed either on NextSeq 500 or Hiseq 2500 illumina sequencers and controlled by the SeqencingAnalysis 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 was from 30 millions to 40 millions.

2018-04-08 | GSE109969 | GEO

NANOG is required for the long-term establishment of avian somatic reprogrammed cells [chicken]

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

NANOG is required for the long-term establishment of avian somatic reprogrammed cells

Project description:NANOG is required for the long-term establishment of avian somatic reprogrammed cells

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NANOG is required for the long-term establishment of avian somatic reprogrammed cells [duck]

Project description:NANOG is required for the long-term establishment of avian somatic reprogrammed cells [duck]

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Acute and long-term effects of mutant p53 in vivo

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

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Establishment of the Primary Avian Gonadal Somatic Cell Lines for Cytogenetic Studies.

Project description:The last decade was marked by a steep rise in avian studies at genomic and cellular levels. Cell lines are important tools for in vitro studies in cell biology and cytogenetics. We developed a simple method of primary somatic cell culture establishment from the ovaries of the great tits (Parus major) and testes of ten Passerine species, characterized the cellular composition of the ovary-derived lines using RT-PCR and immunolocalization of the tissue-specific markers and tested the efficiency of two methods of genetic transformation of the ovary-derived cell line. We found that the ovary-derived cell cultures of the great tit were composed of fibroblasts mainly, but also contained interstitial and granulosa cells. They were cultivated until the 10th passage without any noticeable decrease in their proliferative activity. The testis-derived cell cultures had lower proliferative potential. However, both ovary- and testis-derived cell cultures provided enough material for high quality mitotic metaphase chromosome preparations. The efficiency of its transduction with lentivirus containing a GFP reporter was very low, while electroporation with episomal vectors expressing GFP resulted in a high yield of GFP-positive cells. The proposed method could be used for the generation of high quality material for various cytogenetic and genomic studies.

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Acetyl-CoA metabolism by ACSS2 regulates neuronal histone acetylation and long-term memory

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Establishment of long-term ostracod epidermal culture.

Project description:Primary crustacean cell culture was introduced in the 1960s, but to date limited cell lines have been established. Skogsbergia lerneri is a myodocopid ostracod, which has a body enclosed within a thin, durable, transparent bivalved carapace, through which the eye can see. The epidermal layer lines the inner surface of the carapace and is responsible for carapace synthesis. The purpose of the present study was to develop an in vitro epidermal tissue and cell culture method for S. lerneri. First, an optimal environment for the viability of this epidermal tissue was ascertained, while maintaining its cell proliferative capacity. Next, a microdissection technique to remove the epidermal layer for explant culture was established and finally, a cell dissociation method for epidermal cell culture was determined. Maintenance of sterility, cell viability and proliferation were key throughout these processes. This novel approach for viable S. lerneri epidermal tissue and cell culture augments our understanding of crustacean cell biology and the complex biosynthesis of the ostracod carapace. In addition, these techniques have great potential in the fields of biomaterial manufacture, the military and fisheries, for example, in vitro toxicity testing.

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