Method for efficiently preparing cloned embryos of sheep

Abstract

The invention discloses a method for efficiently preparing high-quality cloned embryos, belonging to the field of embryo transplanting methods. The method provided by the invention comprises the following steps of: agitating and screening to obtain sheep oocytes in the same cell period and transplanting a supplier cell into the sheep oocytes through utilizing microscopy injection; then treating by utilizing a fusion solution and putting the treated embryo into a mature solution to be cultured to form a fused embryo; and chemically activating and then starting and reconstructing embryo cleavage. According to the method provided by the invention, the sheep oocytes which are in the same cell period and have the uniform mature time through an agitating method are acquired, a new method for picking the good sheep oocytes is provided and the mature rate of the sheep oocytes is improved. According to the method provided by the invention, the operation efficiency of nuclear transfer is improved by utilizing a method of simultaneously denucleating through extrusion and carrying out nucleus injection by utilizing the same injection needle. The fusion efficiency is improved to more than 95% by using the fusion solution which is developed by the method provided by the invention, the successful rate of the cloned embryo is greatly improved and the subsequent embryo development is not influenced. The method provided by the invention has the advantages of reducing the loss of the embryos in the process of preparing the cloned embryo and improving the efficiency of preparing the sheep cloned embryos; and the method provided by the invention is a method for efficiently preparing the sheep cloned embryos with good quality and has a wide application prospect.

Description

technical field

[0001] The invention relates to a method for efficiently preparing sheep cloned embryos, belonging to the field of embryo transplantation methods. Background technique

[0002] Currently, there are two main methods for producing mammalian clones: embryo segmentation and nuclear transfer. The cloned sheep "Dolly", as well as various cloned animals bred by scientists from various countries, all adopted the technology of cell nucleus transfer. The so-called nuclear transfer refers to the process of transplanting the nuclei of embryos or adult animals at different developmental stages into oocytes through microinjection and cell fusion methods, reconstituting embryos and making them mature. Different from embryo segmentation technology, nuclear transfer technology, especially serial nuclear transfer technology can produce unlimited genetically identical individuals. Because nuclear transfer is an effective method to produce cloned animals, it is often called an...