Zhao Zhen-Jun, Li Rui-Chang, Cao Heng-Hua, Zhang Quan-Jun, Jiang Man-Xi, Ouyang Ying-Chun, Nan Chang-Long, Lei Zi-Li, Song Xiang-Fen, Sun Qing-Yuan, Chen Da-Yuan
State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Mol Reprod Dev. 2007 Apr;74(4):412-9. doi: 10.1002/mrd.20608.
Interspecies nuclear transfer is an invalulable tool for studying nucleus-cytoplasm interactions; and at the same time, it provides a possible alternative to clone endangered animals whose oocytes are difficult to obtain. In the present study, we investigated the possibility of cloning Tibetan antelope embryos using abattoir-derived caprine oocytes as recipients. Effects of culture conditions, enucleation timing, and donor cell passages on the in vitro development of Tibetan antelope-goat cloned embryos were studied. Maternal to zygotic transition timing of interspecies Tibetan antelope embryos was also investigated using two types of cloned embryos, Tibetan antelope-rabbit and Tibetan antelope-goat embryos. Our results indicate that: (1) goat oocyte is able to reprogram somatic cells of different genus and supports development to blastocyst in vitro. (2) Coculture system supported the development of Tibetan antelope-goat embryos to blastocyst rate stage (4.0%), while CR1aa alone did not. (3) When MII phase enucleated caprine cytoplast and TII phase enucleated caprine cytoplast were used as recipients, the fusion rate and blastocyst rate of hybrid embryos were not statistically different (73.9% vs. 67.4%; 4.0% vs. 1.1%). (4) When donor cells at 3-8 passages were used, 2.9% hybrid embryos developed to blastocysts, while none developed to blastocysts when cells at 10-17 passages were used. (5) There may be a morula-to-blastocyst block for Tibetan antelope-goat, while there may be an 8- to 16-cell block for Tibetan antelope-rabbit embryos.
种间核移植是研究核质相互作用的一项重要工具；同时，它为克隆那些难以获取卵母细胞的濒危动物提供了一种可能的替代方法。在本研究中，我们研究了使用屠宰场来源的山羊卵母细胞作为受体来克隆藏羚羊胚胎的可能性。研究了培养条件、去核时间和供体细胞传代次数对藏羚羊 - 山羊克隆胚胎体外发育的影响。还使用两种克隆胚胎，即藏羚羊 - 兔胚胎和藏羚羊 - 山羊胚胎，研究了种间藏羚羊胚胎从母型向合子型转变的时间。我们的结果表明：（1）山羊卵母细胞能够对不同属的体细胞进行重编程，并支持其在体外发育至囊胚阶段。（2）共培养系统支持藏羚羊 - 山羊胚胎发育至囊胚率阶段（4.0%），而单独使用CR1aa培养基则不能。（3）当使用MII期去核的山羊细胞质体和TII期去核的山羊细胞质体作为受体时，杂交胚胎的融合率和囊胚率没有统计学差异（73.9%对67.4%；4.0%对1.1%）。（4）当使用传代3 - 8次的供体细胞时，2.9%的杂交胚胎发育至囊胚，而使用传代10 - 17次的细胞时，没有胚胎发育至囊胚。（5）藏羚羊 - 山羊胚胎可能存在桑椹胚到囊胚的阻滞，而藏羚羊 - 兔胚胎可能存在8 - 16细胞期的阻滞。
