Improvement of Mouse Cloning from Any Type of Cell by Nuclear Injection.

Somatic cell nuclear transfer (SCNT) technology has become a useful tool for animal cloning, gene manipulation, and genomic reprograming research. The original SCNT was performed using cell fusion between the donor cell and oocyte. This method remains very popular, but we have recently developed an alternative method that relies on nuclear injection rather than cell fusion. The advantages of nuclear injection include a shortened experimental procedure and reduced contamination of donor cytoplasm in the oocyte. In particular, only this method allows us to perform SCNT using dead cells or naked nuclei such as those from cadavers or body wastes. This chapter describes a basic protocol for the production of cloned mice by the nuclear injection method using a piezo-actuated micromanipulator as well as our recent advances in SCNT using noninvasively collected donor cells such as urine-derived somatic cells. This technique will greatly help not only SCNT but also other forms of micromanipulation, including sperm microinjection into oocytes and embryonic stem cell injection into blastocysts.