He Xiao-Ying, Ma Li-Bing, He Xiao-Ning, Si Wan-Tong, Zheng Yue-Mao
School of Life Science and Technology, Inner Mongolia University of Science & Technology, Baotou 014010, China.
Research and Development Center for Tissue Engineering, Xi'an 710048, China.
J Vet Sci. 2016 Jun 30;17(2):145-52. doi: 10.4142/jvs.2016.17.2.145.
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.
先前的研究通过腺病毒介导的端粒酶(hTERT-bMGEs)建立了牛乳腺上皮细胞体外模型。本研究旨在确认hTERT-bMGEs是否为提高转基因表达效率和体细胞核移植(SCNT)的有效靶细胞。为此,使用一个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,未处理的牛乳腺上皮细胞(bMGEs)作为对照组。结果显示,hTERT-bMGEs组的转基因效率和蛋白表达远高于bMGEs组。此外,将非转基因和转基因hTERT-bMGEs用作供体细胞来评估SCNT的效率。第18代和第28代非转基因hTERT-bMGEs组克隆胚胎的卵裂率、囊胚率或孵化囊胚率(分别为82.8%对81.9%、28.6%对24.8%、58.6%对55.3%)与转基因组(80.8%、26.5%和53.4%)之间无显著差异;然而,它们显著高于bMGEs组(71.2%、12.8%和14.8%),(p<0.05)。我们证实hTERT-bMGEs可作为提高体细胞克隆牛胚胎发育的有效靶细胞。