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https://hdl.handle.net/2440/41467
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Type: | Journal article |
Title: | A comparison of two in vitro maturation media for use with adult porcine oocytes for adult somatic cell nuclear transfer |
Author: | Beebe, L. McIlfatrick, S. Grupen, C. Boquest, A. Harrison, S. Faast, R. Ashman, R. Wengle, J. Hamilton, H. Nottle, M. |
Citation: | Cellular Reprogramming, 2007; 9(4):564-570 |
Publisher: | Mary Ann Liebert, Inc. Publishers |
Issue Date: | 2007 |
ISSN: | 1536-2302 1557-7457 |
Statement of Responsibility: | Luke Beebe, Stephen McIlfatrick, Christopher Grupen, Andrew Boquest, Sharon Harrison, Renate Faast, Rodney Ashman, James Wengle, Hamish Hamilton, Mark Nottle |
Abstract: | Two media used to mature adult porcine oocytes for somatic cell nuclear transfer were compared. In the first experiment, parthenogenetic embryos were produced using a maturation medium used by us previously to clone pigs (OMM199) and that described by Kühholzer et al. (2001) to transport oocytes overnight (BOMED). There was no difference in maturation rates between the two different media. However, BOMED medium increased the percentage of parthenogenetic embryos that developed to the blastocyst stage compared with OMM199 (49% vs. 29%, respectively). In a second experiment, BOMED medium increased the percentage of SCNT embryos that developed to the blastocyst stage compared with OMM199 (22% vs. 8%, respectively). The efficiency of our cloning protocol using adult oocytes matured in BOMED medium was then determined by transferring SCNT embryos reconstructed using adult fibroblasts to synchronized recipients. Primary cultures of adult fibroblasts were obtained from two adult male pigs and used for SCNT (passages 2-4). Between 82 and 146 fused couplets were transferred to seven recipients synchronized 1 day behind the embryos. Five recipients (71% pregnancy rate) subsequently farrowed a total of 23 piglets (4.4 average litter size). Overall efficiencies (liveborn/embryos transferred) were 3.2% for all transfers and 4.3% for animals that gave birth. |
Keywords: | Oocytes Cell Nucleus Fibroblasts Animals Swine Culture Media Embryo Transfer Genetic Techniques Cloning, Organism Pregnancy Pregnancy, Animal Female Nuclear Transfer Techniques In Vitro Techniques |
DOI: | 10.1089/clo.2007.0025 |
Published version: | http://dx.doi.org/10.1089/clo.2007.0025 |
Appears in Collections: | Aurora harvest 6 Obstetrics and Gynaecology publications |
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