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

Czernik, Marta; Anzalone, Debora Agata; Palazzese, Luca; Oikawa, Mami; Loi, Pasqualino

doi:10.1387/ijdb.180324mc

Cited by 56 publications

(33 citation statements)

References 76 publications

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“…In turn, an overabundance of the somatic cell-derived agents modulating the epigenetic profile of the donor nucleus may remarkably reduce the concentration and activity of the oocyte's epigenetic factors. Thus, it may diminish the incidence of complete epigenetic reprogramming of transcriptional activity of the somatic cell nucleus in the developing cloned embryo [112,[132][133][134][135][136][137][138][139][140][141].…”

Section: Direct Intraooplasmic Microinjection Of Karyoplastsmentioning

confidence: 99%

Extranuclear Inheritance of Mitochondrial Genome and Epigenetic Reprogrammability of Chromosomal Telomeres in Somatic Cell Cloning of Mammals

Samiec

Skrzyszowska

2021

IJMS

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The effectiveness of somatic cell nuclear transfer (SCNT) in mammals seems to be still characterized by the disappointingly low rates of cloned embryos, fetuses, and progeny generated. These rates are measured in relation to the numbers of nuclear-transferred oocytes and can vary depending on the technique applied to the reconstruction of enucleated oocytes. The SCNT efficiency is also largely affected by the capability of donor nuclei to be epigenetically reprogrammed in a cytoplasm of reconstructed oocytes. The epigenetic reprogrammability of donor nuclei in SCNT-derived embryos appears to be biased, to a great extent, by the extranuclear (cytoplasmic) inheritance of mitochondrial DNA (mtDNA) fractions originating from donor cells. A high frequency of mtDNA heteroplasmy occurrence can lead to disturbances in the intergenomic crosstalk between mitochondrial and nuclear compartments during the early embryogenesis of SCNT-derived embryos. These disturbances can give rise to incorrect and incomplete epigenetic reprogramming of donor nuclei in mammalian cloned embryos. The dwindling reprogrammability of donor nuclei in the blastomeres of SCNT-derived embryos can also be impacted by impaired epigenetic rearrangements within terminal ends of donor cell-descended chromosomes (i.e., telomeres). Therefore, dysfunctions in epigenetic reprogramming of donor nuclei can contribute to the enhanced attrition of telomeres. This accelerates the processes of epigenomic aging and replicative senescence in the cells forming various tissues and organs of cloned fetuses and progeny. For all the above-mentioned reasons, the current paper aims to overview the state of the art in not only molecular mechanisms underlying intergenomic communication between nuclear and mtDNA molecules in cloned embryos but also intrinsic determinants affecting unfaithful epigenetic reprogrammability of telomeres. The latter is related to their abrasion within somatic cell-inherited chromosomes.

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Section: Direct Intraooplasmic Microinjection Of Karyoplastsmentioning

confidence: 99%

Extranuclear Inheritance of Mitochondrial Genome and Epigenetic Reprogrammability of Chromosomal Telomeres in Somatic Cell Cloning of Mammals

Samiec

Skrzyszowska

2021

IJMS

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“…Although SCNT has been successfully used to clone many species of mammals with significant improvements in cloning efficiency in more than 20 years since the birth of Dolly, the proportion of cloned embryos that develop to full term remains very low, greatly limiting the application of SCNT technology (Czernik et al, 2019). To improve the birth of cloned mammals and cloning efficiency, researchers have investigated the effects of donor cell type, oocyte maturation stage, embryo activation method, etc.…”

Section: Low Cloning Efficiency Limits the Applicability Of Scntmentioning

confidence: 99%

Epigenetic Reprogramming During Somatic Cell Nuclear Transfer: Recent Progress and Future Directions

Wang

et al. 2020

Front. Genet.

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Somatic cell nuclear transfer (SCNT) has broad applications but is limited by low cloning efficiency. In this review, we mainly focus on SCNT-mediated epigenetic reprogramming in livestock and also describe mice data for reference. This review presents the factors contributing to low cloning efficiency, demonstrates that incomplete epigenetic reprogramming leads to the low developmental potential of cloned embryos, and further describes the regulation of epigenetic reprogramming by long non-coding RNAs, which is a new research perspective in the field of SCNT-mediated epigenetic reprogramming. In conclusion, this review provides new insights into the epigenetic regulatory mechanism during SCNT-mediated nuclear reprogramming, which could have great implications for improving cloning efficiency.

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“…Attempts are made to overcome these problems to develop and improve the maturation medium from the maturity of the oocytes to the development of embryos in the laboratory by adding supplementation to the ova maturation medium, then fertilization and embryo development ( Mishra et al, 2016 ). Recent reporting of the Red List by International Union for Conservation of Nature (IUCN) has highlighted that twenty-one percent of all mammals are at jeopardy of being extinct globally ( Czernik et al, 2019 , FAO, 2018 ). Various studies have highlighted a scientific conviction that this would allow the cell expansion and reestablishment using the popular and unique somatic cell nuclear transfer (SCNT) technology ( Hildebrandt et al, 2018 , Saragusty et al, 2016 ).…”

Section: Introductionmentioning

confidence: 99%

“…Studies indicate that TSA does induce hyperacetylation of histones and suppresses remethylation of DNA, leading to the un-methylated status of injected nuclei in oocytes ( Mishra et al, 2016 ). Further, this TSA cloning technique led to the first success in cloning the adult Institute of Cancer Research (ICR) mouse, an outbred strain that was directly cloned for the first time in history ( Czernik et al, 2019 ). Beside mouse, some studies showed that TSA treatment resulted in higher pre-implantation embryonic development in pigs ( Wang et al, 2018 ), cattle ( Ding et al, 2008 ), and rabbits ( Zeng et al, 2017 ).…”

Section: Introductionmentioning

confidence: 99%

The in vitro development of cloned sheep embryos treated with Scriptaid and Trichostatin (A)

Al‐Ghadi

Alhimaidi

Iwamoto

et al. 2020

Saudi Journal of Biological Sciences

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Introduction and aims Although, it has been success in the generation of animal clones from somatic cells in various animal species, the information related to nuclear reprogramming of cloned embryos is found to be limited. This study aims to compares the effect of both Scriptaid (SCR) and Trichostatin (A) treatments in improving cloning efficiency, and embryos developmental rate of cloned sheep embryos in vitro . Three groups were formed, i.e., one SCR group, second TSA group, with both treatment concentrations of 5 nM, 50 nM, and 500 nM, respectively, and third were control group with 0 nM. Methods : Ovaries of slaughtered sheep were collected and oocytes were recovered from antral follicles using aspiration method and in vitro maturation of oocytes were done. Then zona dissecting with micropipettes and oocyte enucleation were carried out under the micromanipulator. Later nuclear transfer, cell fusion and activation were done via cell fusion machine. Finally the embryo cultured in incubating chamber at the CO2 incubator up to 9 days. The result : In general the results showed that when the concentration increases the cleavage rate increased. The cleavage rates of the SCNT embryos treated with SCR at different concentrations are closely related to cleavage rate of embryos treated with TSA at same concentration; such as 39.47% for 500 nM TSA, 38.09% for 500 nM SCR; 18.6% for 50 nM TSA, 19.17% for 50 nM SCR, and 22.64% for 5 nM TSA, 17.18% for 5 nM SCR. As for the control group, the cleavage rate of the SCNT embryos cleavage ratewere27.47%., 30% and 30.85% respectively for bothtreatments. While there is a significant difference in TSA treatments at an eight-cell stage at the concentration (5 and 50 nM TSA) compared to the all other cleavage cell stages of (500 nM TSA and control). Also their were a differences between (50 nM of TSA) compared to the (50 nM SCR). Also there were a significant differences between the 16 cell stage at the (500 nM TSA) compared to other treatment (5 nM, 50 nM TSA and control). Regarding the SCR there were a significant difference at 8 cell stage between (5 nM SCR), compared to the other treatment (50 nM, 500 nM SCR and control). Also there were a significant difference at 16 cell stage between (50 nM, and 500 nM SCR), compared to the other treatment (5 nM SCR and control). While in the development of the embryos reach to blastocyst stage the SCR and the control group show a higher rate, in compered to TSA that did not show any development to blastocyst stage. The total SCR treatment showed (3/41 = 7.31%), and the total control showed (4/89 = 4.49%) blastula stage. It concludes that SCR improve the final development blastula stage compared to the TSA treatments that did not improved embryos reach to final developmental blastula stages may be due to spices differences or to the toxicity of TSA, especially at higher concentrations.

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

Cited by 56 publications

References 76 publications

Extranuclear Inheritance of Mitochondrial Genome and Epigenetic Reprogrammability of Chromosomal Telomeres in Somatic Cell Cloning of Mammals

Extranuclear Inheritance of Mitochondrial Genome and Epigenetic Reprogrammability of Chromosomal Telomeres in Somatic Cell Cloning of Mammals

Epigenetic Reprogramming During Somatic Cell Nuclear Transfer: Recent Progress and Future Directions

The in vitro development of cloned sheep embryos treated with Scriptaid and Trichostatin (A)

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