【作者】 董树华；

【导师】 刘俊平； 李光鹏；

【作者基本信息】 内蒙古农业大学 ， 临床兽医学， 2010， 硕士

【摘要】 细胞核移植包括胚胎细胞核移植(Embryo cell nuclear transplantation,简称ECNT)和体细胞核移植( Somatic cell nuclear transfer,简称SCNT),是哺乳动物无性繁殖的主要手段。虽然已经获得了多种存活的克隆动物,但克隆效率的低下仍然是制约该项技术走向产业化的主要瓶颈。除了受体胞质、供体细胞、及克隆操作过程可能影响克隆效率外,围产期受体动物的护理与新生犊牛的护理也极大的影响出生后动物的存活率。到目前为止,尚未有专门对新生克隆犊牛进行系统观察、管理、护理并且提高克隆牛存活率的报道。本研究主要是对牛克隆胚胎的生产,受体牛围产期饲养管理以及新生犊牛出生后的护理等进行了较为系统而详尽的观察与研究。在实验室,主要开展了牛体细胞培养、牛卵母细胞的成熟培养、核移植操作及克隆胚胎的培养等工作。在围产期、犊牛出生过程及出生后进行了详细的观察,主要针对新生犊牛体弱、易感疾病等特点,不同阶段给与不同的护理与治疗措施,尤其是新生克隆犊牛的异物性肺炎、脐带炎及犊牛腹泻等疾病进行了治疗研究。使得体细胞克隆胚妊娠率、出生率以及犊牛出生后存活率大大提高。体细胞克隆胚60d妊娠率为50%,6个月的妊娠率达44%,出生率为33.3%,新生克隆牛犊存活率80%以上。通过我们的系统观察与研究,获得了一套管理、护理克隆犊牛的技术体系,对克隆牛技术产业化,尤其是对转基因克隆牛的生产具有重要的实际意义。更多还原

【Abstract】 Nuclear transfer including embryonic cell nuclear transfer (ECNT) and somatic cell nuclear transfer (SCNT), is a alternative approach in addition to the dominately sexual reproduction. A variety of animal species have been cloned through SCNT technology, the cloning efficiency, however, is extremely low which become the the major bottle-neck obstacle for the economical application of this technology. Although the exact mechanisms affecting the cloning efficiency are not clear, the oocyte cytoplasmic status, donor cells and detailed cloning protocols probably contribute to the extremely low offspring rate. More important, the high death rate of the newborn animals, usually over 50%, makes the cloners and breeders somewhat depression. The recipient cow’s periparturient management and newborn calves’care should have a great impact on the survival rate of the neonatal cloning animals. To our knowledge, this is the first study focused on specific and systematic observations of the cloned calves. During perinatal period, perinatal and the birth of the caves, the recipient cows and the calves were kept an eye on almost 24 hr per day. After transfer of the cloned embryos to the synchronized recipients the pregnancy rate was 50% after 60 days transfer. Sixth month later 44% pregnancy were still maintained, and 33.3% of the recipients delivered healthy calves. Different treatment and management were conducted according to the newborn calf’s appearances and physiological state. A series of physiological indexes such as body temperature, heart rate, respiratory frequency, nose wetness, etc, were measued and careful theropy were taken for the susceptible diseases. Foreign body pneumonia, omphalitis and calf diarrhea are the most susceptible diseases of the newborn cloned calves. With great careful careful treatments and nursings of the recipient cows and the calves the survival rate reached over 80%.Through our careful observations and treatments, a practical protocols for the the managements and nursing of cloned calves were established, which would be beneficial for the industrialization of the cattle cloning and transgenic cloning.更多还原