Developmental competence of HMC™ derived bovine cloned embryos obtained from somatic cell nuclear transfer of adult fibroblasts and granulosa cells
Developmental competence of HMC™ derived bovine cloned embryos obtained from somatic cell nuclear transfer of adult fibroblasts and granulosa cells
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DOI：
10.1262/jrd.17025
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发表时间：
2005-08-01
影响因子：
1.8
通讯作者：
Poehland, R
中科院分区：
文献类型：
--
作者：
Bhojwani, S;Vajta, G;Poehland, R
为了使我们能够在给定的时间内处理大量的卵母细胞，并增加克隆胚胎的生产量，我们尝试了手工克隆（HMC（TM））技术，一种无透明带的牛体细胞核移植方法。我们的目的是研究使用不同类型的体细胞获得的HMC（TM）衍生胚胎的发育能力。共使用6，874个卵丘-卵母细胞复合体，其中第7代或第11代成纤维细胞（分别为第1组和第2组），其从雄性动物制备，或颗粒细胞（第3组）作为核供体。平均卵裂率为65%，卵裂产物的囊胚率仅为2%，> 8细胞胚胎的囊胚率为5%，三组之间没有显著差异。在回收的27个囊胚中，22个囊胚被转移到22个受体，导致两次妊娠。其中一只在第四周后流产，另一只则进展到足月，产下一头雄性小牛。这是欧洲第一次用HMC（TM）技术成功克隆雄性小牛，表明我们实验室成功采用和建立了这项技术，而且这项技术可以成功地产生可存活的胚胎。
To enable us to handle a large number of oocytes at a given time and to have an increased throughput of cloned embryos, we attempted the Handmade cloning (HMC (TM)) technique, a zona-free method of bovine somatic cell nuclear transfer. Our objective was to study the developmental competence of the HMC (TM) derived embryos obtained using different types of somatic cells. A total of 6,874 cumulus-oocyte-complexes were used with either 7th or 11th passage fibroblasts (1st and 2nd groups, respectively), which were prepared from male animals, or granulosa cells (3rd group) as nuclei donors. The average cleavage rate was 65%, accompanied by a blastocyst rate of just 2% for the cleaved products and 5% for the > 8-cell embryos, and there was no significant difference between the three groups. Out of 27 blastocysts recovered, 22 blastocysts were transferred to 22 recipients, resulting in two pregnancies. One pregnancy was lost after the fourth week while the other progressed to full term with the birth of a male calf. This first successful cloning of a male calf with the HMC (TM) technique in Europe indicates the successful adoption and establishment of this technique in our laboratory, and that this technique can be successful in producing viable embryos.