Nuclear Transfer for Cloning Animals

Dinnyés, András; Tian, X. Cindy; Oback, Björn

doi:10.1002/3527600906.mcb.200400139.pub2

Cited by 2 publications

(1 citation statement)

References 191 publications

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“…One of the differences, besides the cell origin is that MEF cells were extensively treated prior to manipulation, whereas cumulus cells were not. For instance, MEF cells were treated with repeated trypsin treatments and kept in culture longer prior to manipulation than cumulus cells (Dinnyes et al, 2004). In accordance with our results, a previous study in the donor cell type of ntESC establishment reported a significantly better rate when freshly isolated cumulus cells were used as donor nuclei compared to cultured tail-tip fibroblast (Wakayama et al, 2005).…”

Section: Zona-free Ntesc Derivationsupporting

confidence: 90%

Generation of Mouse Embryonic Stem Cell Lines from Zona-Free Nuclear Transfer Embryos

Kobolák

Bodó²,

Rungsiwiwut³

et al. 2010

Cellular Reprogramming

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Pluripotent stem cells would have great potential in cell therapies and drug development when genetically matched with the patient; thus, histocompatible cells could be used in transplantation therapy or as a source of patient-specific cells for drug testing. Pluripotent embryonic stem cells (ESCs)-generated via somatic cell nuclear transfer (SCNT) or parthenogenesis (pESC)-are potential sources of histocompatible cells and tissues for transplantation. Earlier studies used the piezoelectric microinjection (PEM) technique for nuclear transfer (NT) in mouse. No specific studies examined zona-free (ZF) NT as an alternative NT method to generate genetically matched ESCs of a nuclear donor. In this study, we compared the efficiency of nuclear transfer-derived ESC (ntESC) line establishment from ZF-NT, ZF-parthenogenetic (PGA), and ZF-fertilized embryos with that of the PEM-NT method. Different nuclei donor cells [cumulus, ESC, and mouse embryonic fibroblast (MEF)] were used and the efficiency of ntESC derivation was investigated, along with their in vitro characterization. The ZF-NT method's efficiency was higher than that of the PEM-NT using cumulus cells. When ESCs and cumulus cells were used as nuclear donor cells, they resulted in significantly higher ZF-NT-derived ntESC line establishment rates compared to MEF cells. In conclusion, the nuclear donor cell type significantly affected the efficiency of ntESC line establishment, and the ZF-NT method was efficient to establish pluripotent ntESC lines.

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Section: Zona-free Ntesc Derivationsupporting

confidence: 90%

Generation of Mouse Embryonic Stem Cell Lines from Zona-Free Nuclear Transfer Embryos

Kobolák

Bodó²,

Rungsiwiwut³

et al. 2010

Cellular Reprogramming

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Quiescence Loosens Epigenetic Constraints in Bovine Somatic Cells and Improves Their Reprogramming into Totipotency

Kallingappa

Turner

Eichenlaub

et al. 2016

Biology of Reproduction

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Reprogramming by nuclear transfer (NT) cloning forces cells to lose their lineage-specific epigenetic marks and reacquire totipotency. This process often produces molecular anomalies that compromise clone development. We hypothesized that quiescence alters the epigenetic status of somatic NT donor cells and elevates their reprogrammability. To test this idea, we compared chromatin composition and cloning efficiency of serum-starved quiescent (G0) fibroblasts versus nonstarved mitotically selected (G1) controls. We show that G0 chromatin contains reduced levels of Polycomb group proteins EED, SUZ12, PHC1, and RING2, as well as histone variant H2A.Z. Using quantitative confocal immunofluorescence microscopy and fluorometric enzyme-linked immunosorbent assay, we further show that G0 induced DNA and histone hypomethylation, specifically at H3K4me3, H3K9me2/3 and H3K27me3, but not H3K9me1. Collectively, these changes resulted in a more relaxed G0 chromatin state. Following NT, G0 donors developed into blastocysts that retained H3K9me3 hypomethylation, both in the inner cell mass and trophectoderm. G0 blastocysts from different cell types and cell lines developed significantly better into adult offspring. In conclusion, serum starvation induced epigenetic changes, specifically hypotrimethylation, that provide a mechanistic correlate for increased somatic cell reprogrammability.

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Nuclear Transfer for Cloning Animals

Cited by 2 publications

References 191 publications

Generation of Mouse Embryonic Stem Cell Lines from Zona-Free Nuclear Transfer Embryos

Generation of Mouse Embryonic Stem Cell Lines from Zona-Free Nuclear Transfer Embryos

Quiescence Loosens Epigenetic Constraints in Bovine Somatic Cells and Improves Their Reprogramming into Totipotency

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