Improvement of mouse cloning using nuclear transfer-derived embryonic stem cells and/or histone deacetylase inhibitor

Wakayama, Sayaka; Wakayama, Teruhiko

doi:10.1387/ijdb.103205sw

Cited by 13 publications

(5 citation statements)

References 80 publications

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“…For this reason, many attempts to modify/improve SCNT efficiency have been reported (Iuso et al, 2013;Czernik et al, 2016). These improvements concerned the technical aspects (Wakayama et al, 2010, Czernik et al, 2016 as well as attempts as bulk or targeted modification of donor nucleus, before or af-et al, 2013). Those authors showed significantly higher blastocyst rates when SCNT was performed without exposure of the oocytes to UV, as compared to the traditional method.…”

Section: Introductionmentioning

confidence: 99%

Somatic cell nuclear transfer: failures, successes and the challenges ahead

Czernik

Anzalone²,

Palazzese³

et al. 2019

Int. J. Dev. Biol.

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Somatic cell nuclear transfer (SCNT) has a broad spectrum of potential applications, including rescue of endangered species, production of transgenic animals, drug production, and regenerative medicine. Unfortunately, the efficiency of SCNT is still disappointingly low. Many factors affecting cloning procedures have been described in several previous reviews; here we review the most effective improvements in SCNT, with a special emphasis on the effect of mitochondrial defects on SCNT embryo/ foetus development, an issue never touched upon before.

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Section: Introductionmentioning

confidence: 99%

Somatic cell nuclear transfer: failures, successes and the challenges ahead

Czernik

Anzalone²,

Palazzese³

et al. 2019

Int. J. Dev. Biol.

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“…Embryonic stem cells derived from cloned embryos (ntESCs) have been established in mouse [14], human [24][25][26], and cattle [27]. In addition, it has been reported that the success rate of cloning was higher when mouse ESCs were used as a source of donor cells compared to more differentiated somatic cells, which are more resistant to epigenetic reprogramming [13,[28][29][30] Contrary to our expectation, the green fluorescent protein following piPSC-nt was expressed in the entire blastocyst rather than just in the ICM. In other words, it was also in the trophoblast.…”

Section: Discussionmentioning

confidence: 99%

Establishment of porcine nuclear transfer-derived embryonic stem cells using induced pluripotent stem cells as donor nuclei

Haraguchi

Dang‐Nguyen

Wells

et al. 2020

Journal of Reproduction and Development

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We investigated whether sequential reprogramming via porcine induced pluripotent stem cells (piPSCs) or exposure to oocyte cytoplasm following nuclear transfer could generate nuclear transfer-derived ESCs (piPSCs-ntESCs). Nuclear transfer embryos were reconstructed with piPSCs possessing a ZsGreen fluorescent marker for expression of exogenous Nanog and Lin28. Reconstructed oocytes developed to morphologically normal 8-cell/morulae (35/93, 37.6%) and blastocysts (12/93, 12.9%). Although most green fluorescent protein-positive blastocysts showed efficient outgrowth (8/10, 80%), none formed primary colonies and all cultures degenerated. Conversely, 15% of fluorescent positive 8-cell/morula stage embryos showed outgrowth (6/40), with three forming primary colonies (7.5%). All three were expanded and maintained as piPSC-ntESC lines. These cell lines expressed stem cell marker genes and proteins. Despite inactivation of one X chromosome, all piPSC-ntESC lines formed teratomas comprising derivatives from all three embryonic germ layers. Strong SSEA1, 3, and 4 expression was detected at the 8-cell/morula stage in embryos reconstructed from both piPSCs and porcine embryonic fibroblasts (PEFs). SSEA3 was notably absent from IVF controls at pre-implantation embryo stages. Finally, we attempted to establish ntESCs from 8-cell/morulae reconstructed with PEFs using the same culture conditions as those for piPSC-ntESC derivation. Although eight primary colonies arose from 107 embryos (7.5%), they all degenerated after the first passage culture. Early and sustained expression of key reprogramming regulatory factors may be critical for pluripotent stem cell derivation to derive piPSC-ntESCs from 8-cell/morula stages, while the expression of SSEAs may be involved in the initial stem cell colony formation phases.

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“…Tail tips derived from mouse ConJ12 (fresh sample) or ConB23 (frozen sample with cell cryopreservation medium "CELLBANKER 1 plus"; Zenoaq) were cultured on a Petri dish for 12-14 d. The cultured tail-tip fibroblasts were used as nuclear donors. After nuclear transfer, the reconstructed oocytes were activated using 5 mM SrCl 2 in Ca-free CZB medium in the presence of 5 μM latrunculin A supplemented with trichostatin A (50 nM) for 9 h. After three washes in CZB, cloned embryos were cultured for 4 d in the same medium, and morula-or blastocyst-stage embryos were used to establish ntES cell lines as described previously (Wakayama et al 2001;Wakayama and Wakayama 2010).…”

Section: Nuclear Transfer and Establishment Of Ntes Cell Linesmentioning

confidence: 99%

Early embryonic mutations reveal dynamics of somatic and germ cell lineages in mice

et al. 2022

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De novo mutations accumulate with zygotic cell divisions. However, the occurrence of these mutations and the way they are inherited by somatic cells and germ cells remain unclear. Here, we present a novel method to reconstruct cell lineages. We identified mosaic mutations in mice using deep whole-genome sequencing and reconstructed embryonic cell lineages based on the variant allele frequencies of the mutations. The reconstructed trees were confirmed using nuclear transfer experiments and the genotyping of approximately 50 offspring of each tree. The most detailed tree had 32 terminal nodes and showed cell divisions from the fertilized egg to germ cell– and somatic cell–specific lineages, indicating at least five independent cell lineages that would be selected as founders of the primordial germ cells. The contributions of each lineage to germ cells and offspring varied widely. At the emergence of the germ cell–specific lineages, 10–15 embryonic mutations had accumulated, suggesting that the pregastrulation mutation rate is 1.0 mutation per mitosis. Subsequent mutation rates were 0.7 for germ cells and 13.2 for tail fibroblasts. Our results show a new framework to assess embryonic lineages; further, we suggest an evolutionary strategy for preserving heterogeneity owing to postzygotic mutations in offspring.

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Improvement of mouse cloning using nuclear transfer-derived embryonic stem cells and/or histone deacetylase inhibitor

Abstract: Nuclear transfer-derived ES (ntES

Cited by 13 publications

References 80 publications

Somatic cell nuclear transfer: failures, successes and the challenges ahead

Somatic cell nuclear transfer: failures, successes and the challenges ahead

Establishment of porcine nuclear transfer-derived embryonic stem cells using induced pluripotent stem cells as donor nuclei

Early embryonic mutations reveal dynamics of somatic and germ cell lineages in mice

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