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.

中文翻译：

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用Scriptaid和Trichostatin（A）处理的克隆绵羊胚胎的体外发育。

介绍和目的

尽管已经成功地从各种动物物种的体细胞产生动物克隆，但是发现与克隆胚胎的核重编程有关的信息是有限的。这项研究旨在比较Scriptaid（SCR）和Trichostatin（A）处理在提高克隆效率和体外克隆绵羊胚胎的胚胎发育率方面的效果。分为三组，即一个SCR组，第二个TSA组，两种治疗浓度分别为5 nM，50 nM和500 nM，第三组为对照组，浓度为0 nM。方法：收集屠宰绵羊的卵巢，采用抽吸法和体外方法从窦房卵泡中回收卵母细胞。卵母细胞成熟。然后用微量移液器解剖透明带并在显微操作器下进行卵母细胞去核。后来的核转移，细胞融合和激活是通过细胞融合机完成的。最后，将胚胎在CO2培养箱的培养箱中培养长达9天。结果：通常，结果表明，当浓度增加时，裂解速率增加。不同浓度的SCR处理的SCNT胚胎的裂解率与相同浓度的TSA处理的胚胎的裂解率密切相关。例如500 nM TSA为39.47％，500 nM SCR为38.09％；50 nM TSA为18.6％，50 nM SCR为19.17％，5 nM TSA为22.64％，5 nM SCR为17.18％。对于对照组，两种处理的SCNT胚胎的切割率分别为27.47％，30％和30.85％。尽管与浓度为5和50 nM TSA的所有其他裂解细胞阶段（500 nM TSA和对照）相比，在八细胞阶段的TSA处理存在显着差异。同样，它们是（50 nM TSA）与（50 nM SCR）之间的差异。与其他治疗（5 nM，50 nM TSA和对照）相比，在（500 nM TSA）的16个细胞阶段之间也存在显着差异。关于SCR，与其他处理（50 nM，500 nM SCR和对照）相比，在8个细胞阶段（5 nM SCR）之间存在显着差异。与其他处理（5 nM SCR和对照）相比，在16个细胞阶段（50 nM和500 nM SCR）之间也存在显着差异。在胚胎发育到胚泡阶段时，SCR和对照组显示出更高的发生率，与TSA相比，胚泡发育没有任何发育。总SCR治疗显示（3/41 = 7.31％），总对照显示（4/89 = 4.49％）囊胚期。