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Improved development of somatic cell cloned bovine embryos by a mammary gland epithelia cells in vitro model

机译:乳腺上皮细胞体外模型改善体细胞克隆牛胚胎的发育

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摘要

Previous studies have established a bovine mammary gland epithelia cells in vitro model by the adenovirus-mediated telomerase (hTERT-bMGEs). The present study was conducted to confirm whether hTERT-bMGEs were effective target cells to improve the efficiency of transgenic expression and somatic cell nuclear transfer (SCNT). To accomplish this, a mammary-specific vector encoding human lysozyme and green fluorescent protein was used to verify the transgenic efficiency of hTERT-bMGEs, and untreated bovine mammary gland epithelial cells (bMGEs) were used as a control group. The results showed that the hTERT-bMGEs group had much higher transgenic efficiency and protein expression than the bMGEs group. Furthermore, the nontransgenic and transgenic hTERT-bMGEs were used as donor cells to evaluate the efficiency of SCNT. There were no significant differences in rates of cleavage or blastocysts or hatched blastocysts of cloned embryos from nontransgenic hTERT-bMGEs at passage 18 and 28 groups (82.8% vs. 81.9%, 28.6% vs. 24.8%, 58.6% vs. 55.3%, respectively) and the transgenic group (80.8%, 26.5% and 53.4%); however, they were significantly higher than the bMGEs group (71.2%, 12.8% and 14.8%), (p < 0.05). We confirmed that hTERT-bMGEs could serve as effective target cells for improving development of somatic cell cloned cattle embryos.
机译:先前的研究已通过腺病毒介导的端粒酶(hTERT-bMGEs)建立了体外的牛乳腺上皮细胞模型。进行本研究以证实hTERT-bMGEs是否是有效的靶细胞,以提高转基因表达和体细胞核转移(SCNT)的效率。为此,使用编码人溶菌酶和绿色荧光蛋白的乳腺特异性载体来验证hTERT-bMGEs的转基因效率,并将未经处理的牛乳腺上皮细胞(bMGEs)用作对照组。结果显示,hTERT-bMGEs组比bMGEs组具有更高的转基因效率和蛋白质表达。此外,将非转基因和转基因的hTERT-bMGEs用作供体细胞以评估SCNT的效率。在第18和28代时,来自非转基因hTERT-bMGEs的克隆胚胎的卵裂,囊胚或孵出的囊胚率没有显着差异(82.8%vs. 81.9%,28.6%vs 24.8%,58.6%vs. 55.3%,分别)和转基因组(分别为80.8%,26.5%和53.4%);但是,它们显着高于bMGEs组(71.2%,12.8%和14.8%),(p <0.05)。我们证实,hTERT-bMGEs可以作为有效的靶细胞,用于改善体细胞克隆牛胚胎的发育。

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