牛脂肪组织来源干细胞胚胎与体外和孤雌胚胎的表观遗传修饰基因比较

M. Salehi, B. Abouhamzeh, A. Hosseini, Z. Zare, A. Bakhtari
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引用次数: 1

摘要

目的考虑到未分化间充质干细胞作为供体细胞需要较少的表观遗传重编程,评估使用低水平DNMTs和hdac表达的牛脂肪组织源性干细胞(BASCs)的可能性。材料与方法在本实验研究中,我们检测了体外受精(IVF)、孤雌生殖激活(PA)和体细胞核移植(SCNT)各组着床前发育两个阶段赖氨酸9 (H3K9ac)和POU5F1(也称为OCT4)组DNA甲基转移酶(DNMT1、DNMT3A和DNMT3B)和组蛋白去乙酰化酶(HDAC1-3)等表观遗传修饰因子的基因表达,以及组蛋白H3乙酰化蛋白水平。结果体外受精胚胎的发育能力高于SCNT胚胎(P<0.05)。在PA和SCNT组中,DNMT1、HDAC2和HDAC3 mRNA过表达(P<0.05), H3K9ac和POU5F1蛋白水平在6-8细胞和囊胚期与IVF相比降低(P<0.05)。DNMT1 mRNA的表达在两个发育阶段均<0.05(囊胚期除外)。结论来源于BASCs的SCNT胚胎在胚胎早期发育过程中经历了大量的核重编程。PA和SCNT囊胚的比较表明,HDAC1和DNMT1可能与牛胚胎的发育能力差异有关。
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Comparison of Epigenetic Modifier Genes in Bovine Adipose Tissue-Derived Stem Cell Based Embryos, as Donors, with In Vitro and Parthenogenesis Embryos
Objective Regarding that undifferentiated mesenchymal stem cells, as donor cells, require less epigenetic reprogramming, possibility of using bovine adipose tissue-derived stem cells (BASCs) with low level of DNMTs and HDACs expression was evaluated. Materials and Methods In this experimental study, we examined gene expression of epigenetic modifiers including DNA methyltransferases (DNMT1, DNMT3A and DNMT3B) and histone deacetylases (HDAC1-3), as well as protein levels of histone H3 acetylation at lysine 9 (H3K9ac) and POU5F1 (also known as OCT4) at two stages of preimplantation development among in vitro fertilization (IVF), parthenogenetic activation (PA) and somatic cell nuclear transfer (SCNT) groups. Results The results revealed that developmental competence of IVF embryos was higher than SCNT embryos (P<0.05). In the PA and SCNT groups, DNMT1, HDAC2 and HDAC3 mRNA were overexpressed (P<0.05), and proteins levels of H3K9ac and POU5F1 were reduced at 6-8 cells and blastocyst stages compared to IVF (P<0.05). The mRNA expression of DNMT1 an<0.05) in both developmental stages (except HDAC1 in blastocyst stage). Conclusion The SCNT embryos derived from BASCs have endured considerable nuclear reprogramming during early embryo development. Comparison of PA and SCNT blastocysts demonstrated that HDAC1 and DNMT1 may attribute to developmental competence variability of bovine embryos.
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