title: Nuclear transfer in the mouse.
abstract: Nuclear transfer (NT) technique provides a powerful experimental tool to study the mechanisms of reprogramming processes and to derive NT-embryonic stem (ntES) cells from living or frozen animals. The Piezo-driven direct microinjection NT method has proved to be a valid technique to clone mice and other species. In addition, this method has been broadly used as a versatile tool for many fields of mouse micromanipulation. This chapter describes the "one step method" protocol of nuclear transfer in mouse, which combines injection of a donor cell nucleus and enucleation of MII metaphase in a single manipulation procedure. This protocol describes the isolation and collection of oocytes, treatment of donor cells, visualization of spindle-chromosomal complex, direct injection and enucleation, activation of reconstructed embryos and their in vitro culture and transfer into pseudopregnant mice.
authors.affiliation: INRA, UMR1198 Biologie du Développement et Reproduction, Jouy-en-Josas, 78350, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Brochard
authors.forename: Zichuan
authors.initials: Z
authors.lastname: Liu
mesh_terms: D000818:Animals; D046149:Embryo Culture Techniques; D004624:Embryo Transfer / Q000379:methods*; D005260:Female; D005347:Fibroblasts / Q000166:cytology; D051379:Mice; D008815:Mice, Inbred Strains; D008845:Microinjections; D008846:Micromanipulation / Q000295:instrumentation / Q000379:methods; D053652:Nuclear Transfer Techniques; D054315:Oocyte Retrieval / Q000379:methods*; D011247:Pregnancy