Oh Hyun Ju, Choi Jin, Kim Min Jung, Kim Geon A, Jo Young Kwang, Choi Yoo Bin, Lee Byeong Chun
Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Seoul National University, Seoul, Korea.
Anim Sci J. 2016 Jan;87(1):21-6. doi: 10.1111/asj.12402. Epub 2015 Aug 7.
The objective of the present study was to compare the efficiency of two oocyte activation culture media to produce cloned dogs from an elite rescue dog and to analyze their behavioral tendencies. In somatic cell nuclear transfer procedure, fused couplets were activated by calcium ionophore treatment for 4 min, cultured in two media: modified synthetic oviduct fluid (mSOF) with 1.9 mmol/L 6-dimethylaminopyridine (DMAP) (SOF-DMAP) or porcine zygote medium (PZM-5) with 1.9 mmol/L DMAP (PZM-DMAP) for 4 h, and then were transferred into recipients. After embryo transfer, pregnancy was detected in one out of three surrogate mothers that received cloned embryos from the PZM-DMAP group (33.3%), and one pregnancy (25%) was detected in four surrogate mothers receiving cloned embryos from the SOF-DMAP group. Each pregnant dog gave birth to one healthy cloned puppy by cesarean section. We conducted the puppy aptitude test with two cloned puppies; the two cloned puppies were classified as the same type, accepting humans and leaders easily. The present study indicated that the type of medium used in 6-DMAP culture did not increase in cloning efficiency and dogs cloned using donor cells derived from one elite dog have similar behavioral tendencies.
本研究的目的是比较两种卵母细胞激活培养基从一只优秀救援犬培育克隆犬的效率,并分析它们的行为倾向。在体细胞核移植过程中,融合的细胞对经钙离子载体处理4分钟进行激活,然后在两种培养基中培养4小时:含有1.9 mmol/L 6-二甲基氨基吡啶(DMAP)的改良合成输卵管液(mSOF)(SOF-DMAP)或含有1.9 mmol/L DMAP的猪合子培养基(PZM-5)(PZM-DMAP),之后转移至受体动物体内。胚胎移植后,在从PZM-DMAP组接受克隆胚胎的三只代孕母犬中有一只检测到怀孕(33.3%),在从SOF-DMAP组接受克隆胚胎的四只代孕母犬中有一只检测到怀孕(25%)。每只怀孕的母犬通过剖宫产产下一只健康的克隆幼犬。我们对两只克隆幼犬进行了幼犬资质测试;这两只克隆幼犬被归为同一类型,很容易接受人类和领导者。本研究表明,6-DMAP培养中使用的培养基类型并未提高克隆效率,并且使用来自一只优秀犬的供体细胞克隆的犬具有相似的行为倾向。