Effect of donor cell type and gender on the efficiency of in vitro sheep somatic cell cloning
Effect of donor cell type and gender on the efficiency of in vitro sheep somatic cell cloning
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DOI：
10.1016/j.smallrumres.2008.06.004
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发表时间：
2008-08-01
影响因子：
1.8
通讯作者：
Nasr-Esfahani, M. H.
中科院分区：
文献类型：
--
作者：
Hosseini, S. M.;Moulavi, F.;Nasr-Esfahani, M. H.
尽管在使用不同体细胞核克隆哺乳动物方面取得了显着进展，但关于哺乳动物体细胞核移植供体细胞来源（类型和/或性别）的选择的信息很少。本研究旨在研究使用不同细胞来源的供体细胞核进行体外绵羊体细胞核移植的效率。供体成纤维细胞系是从雄性和雌性成年绵羊的活检中产生的，而卵丘细胞系是从同一只母羊中收获的。为了使细胞周期同步化，将供体细胞培养至汇合，然后在血清饥饿培养基中培养4-5天。使用体外成熟 22 小时的屠宰场来源的绵羊卵母细胞进行去核。使用直接全细胞注射程序重建去核卵母细胞，并将活化的重建胚胎在SOF培养基中培养8天。在整个研究中，与卵丘细胞相比，核移植过程中卵母细胞裂解和体外胚胎发育过程中碎裂变性的比例显着更高，这可能是由于与卵丘细胞相比，成纤维细胞的细胞尺寸较大且卵母细胞兼容性较差。关于体外克隆胚胎发育率，记录的唯一显着差异是在 5-8 细胞阶段的卵丘和雄性成纤维细胞与雌性成纤维细胞之间（分别为 70.7% 和 82.6% 对 56.5%），以及在 8-16 细胞状态的雄性成纤维细胞与卵丘细胞之间（分别为 60.9% 对 33.8%）。卵丘、男性和女性成纤维细胞供体细胞的囊胚形成率没有显着差异（分别为 19.5%、17.4% 和 15.2%）。由于三个SCNT组之间体外胚胎发育的总体比率没有差异，因此可以得出结论，供体细胞的类型和护栏对SCNT绵羊胚胎体外生产的总体效率没有显着影响。 (C) 2008 Elsevier B.V. 保留所有权利。
Despite remarkable progress in the cloning of mammals Using different somatic nuclei, there is little information available regarding the choice of donor cell origin (type and/or gender) for mammalian somatic cell nuclear transfer. This study was designed to investigate the efficiency of in vitro sheep somatic cell nuclear transfer using donor nuclei derived from different cell origins. Donor fibroblast cell lines were produced from biopsies taken from a male and a female adult sheep and cumulus cell lines were harvested from the same ewe. For cell cycle synchronization, donor cells were cultured to confluence and then subjected to a serum-starved medium for a period of 4-5 days. Abattoir-derived sheep oocytes matured in vitro for 22 h were used for enucleation. A direct, whole cell injection procedure was used for reconstitution of the enucleated oocytes and the activated reconstructed embryos were cultured in SOF medium for 8 days. Throughout the study, the ratio of oocyte lysis during nuclear transfer and fragmentation-degeneration during in vitro embryo development were significantly higher in the reconstructed fibroblast vs. the cumulus cells-possibly due to the larger cell size and less oocyte-compatibility of the fibroblast, compared to the cumulus cells. Regarding the rate of in vitro cloned embryo development, the only significant difference recorded was between the cumulus and male fibroblast vs. the female fibroblast cells at the 5-8 cell stage (70.7% and 82.6% vs. 56.5%, respectively) and between the male fibroblast with cumulus cells at the 8-16 cell state (60.9% vs. 33.8%, respectively). The blastocyst formation rates were not significantly different between the cumulus, male and female fibroblast donor cells (19.5%, 17.4% and 15.2%, respectively). As the overall ratios of in vitro embryo development were not different between the three SCNT groups, it could be concluded that neither the type of the donor cell nor fender has a significant effect on the overall efficiency of in vitro production of SCNT sheep embryo production. (C) 2008 Elsevier B.V. All rights reserved.