Comet assay on thawed embryos: An optimized technique to evaluate DNA damage in mouse embryos

Rolland, Laura; Courbière, Blandine; Tassistro, Virginie; Sansoni, Amandine; Orsière, Thierry; Liu, Wei; Giorgio, Carole Di; Perrin, Jeanne

doi:10.1016/j.tiv.2017.07.010

Cited by 5 publications

(4 citation statements)

References 51 publications

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“…The reference materials must be stable during storage and shipment to the user and in sufficient supply to distribute to the various laboratories participating in a coordinated study. Important cryopreservation procedures have recently been described [ 16 , 94 ]. One particular concern with chemical methods used to produce DNA damage is that residual amounts of the genotoxic agent may continue to cause further damage, even after characterization and during storage.…”

Section: Discussionmentioning

confidence: 99%

Standards for Quantitative Measurement of DNA Damage in Mammalian Cells

Atha

Reipa

2023

IJMS

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As the potential applications of DNA diagnostics continue to expand, there is a need for improved methods and standards for DNA analysis. This report describes several methods that could be considered for the production of reference materials for the quantitative measurement of DNA damage in mammalian cells. With the focus on DNA strand breaks, potentially useful methods for assessing DNA damage in mammalian cells are reviewed. The advantages and limitations of each method, as well as additional concerns with respect to reference material development, are also discussed. In conclusion, we outline strategies for developing candidate DNA damage reference materials that could be adopted by research laboratories in a wide variety of applications.

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Section: Discussionmentioning

confidence: 99%

Standards for Quantitative Measurement of DNA Damage in Mammalian Cells

Atha

Reipa

2023

IJMS

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“…[26] For performing this test, after sperm cell lysis, uses detergents such as sodium dodecyl sulfate (SDS) or mercaptoethanol to move and detach DNA proteins, and then they are placed on agarose in an electric playing field. [27] During electrophoresis, fragmented DNA that are characterized by fluorescence colors are separated from the chromatin and move in the direction of the anode. Sperms with fragmented DNA have a longer tail in a comet form and look like a tear drop that is due to the migration and stretching of short DNA fragments.…”

Section: Methods Of Investigation Of Sperm Dna Damagementioning

confidence: 99%

Cytochemical tests to investigate sperm DNA damage: Assessment and review

Pourmasumi

Nazari

Fagheirelahee

et al. 2019

J Family Med Prim Care

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Male factor infertility has been diagnosed as the cause of infertility in about 20% of infertile couples. Sperm analysis is the most common method for diagnosing infertility in a laboratory. However, approximately 15% of infertile men have a normal sperm analysis. Therefore, the result of a routine sperm analysis often cannot be a definitive diagnosis for male factor infertility. Also, approximately 8% of infertile men with normal sperm parameters have high levels of abnormal sperm DNA. This indicates the role of the integrity of sperm DNA in male infertility. Here, we review the current tests available to evaluate the sperm DNA integrity along with their benefits and limitations.

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“…Although in the reproduction field, this technique is mainly used to assess sperm quality and sometimes in oocytes (Chang et al, 2019;Haddock et al, 2020), it has also been adapted to study early embryos, even though the number of studies are very limited and comprise different species. One of its applications is the evaluation of the persistence of induced damage from maternal germ cells in early embryos (Rolland et al, 2017). However, an important limitation of this technique is that it cannot distinguish between DNA fragmentation coming from polar body disintegration (Tatemoto et al, 2000) and DNA fragmentation induced through apoptosis in embryonic cells (Fabian et al, 2003).…”

Section: Assessment Of Embryo Quality Through Senescence and Apoptosimentioning

confidence: 99%

Senescence and Apoptosis During in vitro Embryo Development in a Bovine Model

Ramos‐Ibeas

Gimeno

Cañón-Beltrán

et al. 2020

Front. Cell Dev. Biol.

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According to the World Health Organization, infertility affects up to 14% of couples under reproductive age, leading to an exponential rise in the use of assisted reproduction as a route for conceiving a baby. In the same way, thousands of embryos are produced in cattle and other farm animals annually, leading to increased numbers of individuals born. All reproductive manipulations entail deviations of natural phenotypes and genotypes, with in vitro embryo technologies perhaps showing the biggest effects, although these alterations are still emerging. Most of these indications have been provided by animal models, in particular the bovine species, due to its similarities to human early embryo development. Oocytes and embryos are highly sensitive to environmental stress in vivo and in vitro. Thus, during in vitro culture, a number of stressful conditions affect embryonic quality and viability, inducing subfertility and/or long-term consequences that may reach the offspring. A high proportion of the embryos produced in vitro are arrested at a species-specific stage of development during the first cell divisions. These arrested embryos do not show signs of programmed cell death during early cleavage stages. Instead, defective in vitro produced embryos would enter a permanent cell cycle arrest compatible with cellular senescence, in which they show active metabolism and high reactive oxygen species levels. Later in development, mainly during the morula and blastocyst stages, apoptosis would mediate the elimination of certain cells, accomplishing both a physiological role in to balancing cell proliferation and death, and a pathological role preventing the transmission of damaged cells with an altered genome. The latter would acquire relevant importance in in vitro produced embryos that are submitted to stressful environmental stimuli. In this article, we review the mechanisms mediating apoptosis and senescence during early embryo development, with a focus on in vitro produced bovine embryos. Additionally, we shed light on the protective role of senescence and apoptosis to ensure that unhealthy cells and early embryos do not progress in development, avoiding long-term detrimental effects.

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Comet assay on thawed embryos: An optimized technique to evaluate DNA damage in mouse embryos

Cited by 5 publications

References 51 publications

Standards for Quantitative Measurement of DNA Damage in Mammalian Cells

Standards for Quantitative Measurement of DNA Damage in Mammalian Cells

Cytochemical tests to investigate sperm DNA damage: Assessment and review

Senescence and Apoptosis During in vitro Embryo Development in a Bovine Model

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