title: Production of cloned mice by nuclear transfer of cumulus cells.
abstract: BACKGROUND
  Over the past several years, mammals have been successfully cloned by either the splitting of an early stage embryo or nuclear transfer of adult somatic cells (NT) into oocytes. Although it has been 15 years since the generation of the first cloned mammals from somatic cells by NT, the success rate for producing live offspring by this technique is low regardless of the cell type and animal species used. However, these techniques have the potential to be important tools for future research in basic biology. In the present study, we described our experiences in producing successfully cloned mouse using NT method and piezo-actuated micromanipulator.
  
  
  METHODS
  B6D2F1 mice, 8-12 weeks old, were superovulated with injections of 5 IU of pregnant mare serum gonadotropin and 5 IU of human chorionic gonadotropin administered 48 hr apart. Enucleation and donor nuclei cumulus cell injection were performed with a piezo-actuated micromanipulator after which activation and trichostatin A treatment were used for reconstructed oocytes. Two-cell stage cloned embryos that developed in the mWM medium were transferred into the oviducts of pseudopregnant NMRI mice.
  
  
  RESULTS
  Of 367 oocytes collected, 131 (69%) developed into 2-cell stage embryos. Of these, 5 (1%) live pups were successfully delivered. We used NMRI foster mother to raise the pups by lactation. One adult cloned mouse was mated, after which she delivered and raised normal offspring.
  
  
  CONCLUSIONS
  For mouse cloning, the present study also successfully tested the capability of somatic cell nuclear transfer SCNT using a piezo unit.
authors.affiliation: Department of Biological Sciences and Biotechnology, School of Science, University of Kurdistan, Sanandaj, Iran ; These authors equally contributed to this work.
authors.forename: Soleiman
authors.initials: S
authors.lastname: Kurd
authors.forename: Mohammad Ali
authors.initials: MA
authors.lastname: Zarei
authors.forename: Fardin
authors.initials: F
authors.lastname: Fathi
authors.forename: Tayyeb
authors.initials: T
authors.lastname: Ghadimi
authors.forename: Mohammad Saeed
authors.initials: MS
authors.lastname: Hakhamaneshi
authors.forename: Ali
authors.initials: A
authors.lastname: Jalili
keywords: Mouse; Nuclear transfer techniques; Trichostatin A
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