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

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...

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Autores principales: Al-Ghadi, Muath Q, Alhimaidi, Ahmad R, Iwamoto, Daisaku, AL-Mutary, Mohsen G., Ammari, Aiman A, Saeki, Kazuhiro O, Aleissa, Mohammed S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7451688/
https://www.ncbi.nlm.nih.gov/pubmed/32884408
http://dx.doi.org/10.1016/j.sjbs.2020.04.039
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author Al-Ghadi, Muath Q
Alhimaidi, Ahmad R
Iwamoto, Daisaku
AL-Mutary, Mohsen G.
Ammari, Aiman A
Saeki, Kazuhiro O
Aleissa, Mohammed S.
author_facet Al-Ghadi, Muath Q
Alhimaidi, Ahmad R
Iwamoto, Daisaku
AL-Mutary, Mohsen G.
Ammari, Aiman A
Saeki, Kazuhiro O
Aleissa, Mohammed S.
author_sort Al-Ghadi, Muath Q
collection PubMed
description 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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spelling pubmed-74516882020-09-02 The in vitro development of cloned sheep embryos treated with Scriptaid and Trichostatin (A) Al-Ghadi, Muath Q Alhimaidi, Ahmad R Iwamoto, Daisaku AL-Mutary, Mohsen G. Ammari, Aiman A Saeki, Kazuhiro O Aleissa, Mohammed S. Saudi J Biol Sci Article 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. Elsevier 2020-09 2020-04-29 /pmc/articles/PMC7451688/ /pubmed/32884408 http://dx.doi.org/10.1016/j.sjbs.2020.04.039 Text en © 2020 The Authors http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Al-Ghadi, Muath Q
Alhimaidi, Ahmad R
Iwamoto, Daisaku
AL-Mutary, Mohsen G.
Ammari, Aiman A
Saeki, Kazuhiro O
Aleissa, Mohammed S.
The in vitro development of cloned sheep embryos treated with Scriptaid and Trichostatin (A)
title The in vitro development of cloned sheep embryos treated with Scriptaid and Trichostatin (A)
title_full The in vitro development of cloned sheep embryos treated with Scriptaid and Trichostatin (A)
title_fullStr The in vitro development of cloned sheep embryos treated with Scriptaid and Trichostatin (A)
title_full_unstemmed The in vitro development of cloned sheep embryos treated with Scriptaid and Trichostatin (A)
title_short The in vitro development of cloned sheep embryos treated with Scriptaid and Trichostatin (A)
title_sort in vitro development of cloned sheep embryos treated with scriptaid and trichostatin (a)
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7451688/
https://www.ncbi.nlm.nih.gov/pubmed/32884408
http://dx.doi.org/10.1016/j.sjbs.2020.04.039
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