Xiong X, Li J, Wang L, Zhong J, Zi X, Wang Y
College of Life Science and Technology, Southwest University for Nationalities, Chengdu, Sichuan, China.
Reprod Domest Anim. 2014 Feb;49(1):126-33. doi: 10.1111/rda.12240. Epub 2013 Sep 17.
With an aim to improve the efficiency of yak-bovine interspecies somatic cell nuclear transfer (iSCNT), this study investigated the effect of different culture systems on the development, quality and gene expression profile of yak-bovine iSCNT embryo. Reconstructed embryos were cultured in modified synthetic oviductal fluid (mSOF) or relative defined culture medium (RDCM) with 5% or 20% oxygen tension. Relative mRNA abundance of Oct-4, IFNT, IGF-2, Bax, GPX-1, SOD-1, CAT and GSS was analysed in blastocysts with qRT-PCR. The blastocyst formation rate in RDCM under 5% oxygen tension was significantly higher than that under 20% oxygen tension (P < 0.05). The total cell number of blastocyst derived from RDCM with 20% oxygen tension was lower than that of other groups, whereas the group of RDCM with 5% oxygen tension showed a beneficial effect on apoptosis index and tolerance to cryopreservation (P < 0.05). However, under the same oxygen tension, the mRNA abundance of IFNT of RDCM groups was higher than that of the mSOF groups. In addition, high oxygen tension during in vitro culture (IVC) with RDCM significantly increases the mRNA expression of oxidative stress-related genes (GPX-1, SOD-1, CAT and GSS) (P < 0.05). 3, 4-Dihydroxyflavone (DHF) during high oxygen tension was able to improve the cloned blastocyst formation rate in RDCM (P < 0.05). These results for the first time showed that low oxygen tension and RDCM could improve the developmental competence and quality and alleviate the oxidative stress for yak-bovine iSCNT embryo during IVC.
为提高牦牛 - 牛种间体细胞核移植(iSCNT)的效率,本研究探讨了不同培养体系对牦牛 - 牛iSCNT胚胎发育、质量和基因表达谱的影响。将重构胚胎培养于改良的合成输卵管液(mSOF)或相对限定培养基(RDCM)中,氧张力分别为5%或20%。采用qRT-PCR分析囊胚中Oct-4、IFNT、IGF-2、Bax、GPX-1、SOD-1、CAT和GSS的相对mRNA丰度。5%氧张力下RDCM中的囊胚形成率显著高于20%氧张力下的(P < 0.05)。20%氧张力下RDCM来源的囊胚总细胞数低于其他组,而5%氧张力的RDCM组对凋亡指数和冷冻耐受性有有益影响(P < 0.05)。然而,在相同氧张力下,RDCM组的IFNT mRNA丰度高于mSOF组。此外,在RDCM中进行体外培养(IVC)时高氧张力显著增加了氧化应激相关基因(GPX-1、SOD-1、CAT和GSS)的mRNA表达(P < 0.05)。在高氧张力期间添加3,4 - 二羟基黄酮(DHF)能够提高RDCM中的克隆囊胚形成率(P < 0.05)。这些结果首次表明,低氧张力和RDCM可提高牦牛 - 牛iSCNT胚胎在IVC期间的发育能力和质量,并减轻氧化应激。