Birth of Cloned Pigs from Zona-Free Nuclear Transfer Blastocysts DevelopedIn Vitrobefore Transfer
- 1 December 2006
- journal article
- Published by Mary Ann Liebert Inc in Cloning and Stem Cells
- Vol. 8 (4), 283-293
- https://doi.org/10.1089/clo.2006.8.283
Abstract
The objective of this work was to obtain cloned pig offspring by uterine transfer of blastocysts produced by zona-free manipulation. We started by defining the most suitable culture media for growing pig nuclear transfer embryos produced by zona-free micromanipulation comparing NCSU-23aa with Synthetic Oviduct Fluid (SOFaa) and with in vivo culture in the sheep oviduct. We found that parthenogenetic development to day 7 blastocyst in NCSU-23aa and sheep oviduct was significantly superior as compared to SOFaa (61.8%, 64% and 42.4 respectively) although blastocyst cell number was higher in the latter. Interestingly, when we compared the two media for the culture of nuclear transfer (NT) embryos derived from 3 different donor cell lines, we observed lower rates of development with NCSU-23aa (from 24.5% to 32.4%) while with SOFaa the development was significantly higher for two donor cell lines as compared to the third (44.4%, 48.9% and 20.6% respectively). A total of 244 blastocysts grown in SOFaa were transferred in four synchronized sows on day 5 or 6 of development. Two recipients farrowed 6 and 8 piglets corresponding to an efficiency of development to term of 8% and 16% of the transferred embryos respectively. Eleven pigs are now 10 month of age and those that have reached puberty have been proven to be fertile. Finally, this is the first report on the production of cloned pigs derived from the transfer of NT embryos at the blastocyst stage.Keywords
This publication has 50 references indexed in Scilit:
- Birth of piglets after transferring of in vitro-produced embryos pre-matured with R-roscovitineReproduction, 2005
- Embryos Derived from Porcine Skin-Derived Stem Cells Exhibit Enhanced Preimplantation Development1Biology of Reproduction, 2004
- Production of Piglets Derived from In Vitro-Produced Blastocysts Fertilized and Cultured in Chemically Defined Media: Effects of Theophylline, Adenosine, and Cysteine During In Vitro Fertilization1Biology of Reproduction, 2003
- Introduction to Cloning by Nuclear TransplantationCloning and Stem Cells, 2003
- Relationship between donor animal age, follicular fluid steroid content and oocyte developmental competence in the pigReproduction, Fertility and Development, 2003
- Comparison of Microinjection (Piezo-Electric) and Cell Fusion for Nuclear Transfer Success with Different Cell Types in CattleCloning and Stem Cells, 2002
- Kinetics of Oocyte Maturation and Subsequent Development of IVF, Parthenogenetic, and NT Bovine Embryos after Meiotic Inhibition with RoscovitineCloning and Stem Cells, 2002
- Application of the Zona-Free Manipulation Technique to Porcine Somatic Nuclear TransferCloning and Stem Cells, 2001
- Significance of the Number of Embryonic Cells and the State of the Zona Pellucida for Hatching of Mouse Blastocysts In Vitro Versus In VivoBiology of Reproduction, 2000
- Chronological and cytological details of fertilization and early embryonic development in the domestic pig, Sus scrofaThe Anatomical Record, 1974