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Titlebook: Nuclear Reprogramming; Methods and Protocol Nathalie Beaujean,Hélène Jammes,Alice Jouneau Book 2015Latest edition Springer Science+Business

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Nuclear Transfer in Ruminants,t animals (Willadsen, Nature 320(6057):63–65, 1986; Prather et al., Biol Reprod 37(4):859–866, 1987; Wilmut et al., Nature 385(6619):810–813, 1997). Since then, cloning has provided us with a vast amount of knowledge and information on the reprogramming ability of somatic cells to different cell typ
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Nuclear Transfer and Transgenesis in the Pig,and for xenotransplantation. Up to now, SCNT is the main way to generate gene-targeted pigs, since germ line-competent pluripotent stem cells are not available for this species. In this chapter, we introduce our routine workflow for the production of genetically engineered pigs, especially focused o
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,Embryonic Stem Cell–Somatic Cell Fusion and Postfusion Enucleation,ties to pluripotent cells. It has also been shown that transcriptional changes can occur in a heterokaryon, without nuclear hybridization. However it is unclear whether these changes can be sustained following removal of the dominant ES nucleus. In this chapter, methods are described for the cell fu
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Analysis of Nuclear Reprogramming Following Nuclear Transfer to , Oocyte, nuclear transfer no cell division occurs and no new cell types are generated. However, the transplanted nuclei undergo extensive transcriptional reprogramming..Here, it is first explained how to carry out transplantation of multiple mammalian cell nuclei to . oocytes. It is then described how to pe
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Assessing the Quality of Donor Cells: Karyotyping Methods,nced by the biological material used (oocyte and donor cell quality). Hence, it is crucial to check the normality of the donor cell’s karyotype. Numerical and structural chromosome abnormalities are detected by cytogenetic analysis at minimum using G-banding to identify the chromosomes. Here, we des
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Live Embryo Imaging to Follow Cell Cycle and Chromosomes Stability After Nuclear Transfer,bryos consider only embryos at predefined key stages (e.g., morula or blastocyst), that is, after the bulk of reprogramming has taken place. These retrospective approaches are of limited use to elucidate mechanisms of reprogramming and to predict developmental success. Observing cloned embryo develo
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Analysis of Nucleolar Morphology and Protein Localization as an Indicator of Nuclear Reprogramming,rence of the reprogramming. This phenomenon is most pronounced when differentiated cells are reprogrammed to totipotency when they are submitted to cloning by somatic cell nuclear transfer. However, when cells are reprogrammed by less fundamental means, as for example treatment by Xenopus extract or
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