Please use this identifier to cite or link to this item: http://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: Cloning and Stem Cells, 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
RMID: 0020074104
DOI: 10.1089/clo.2007.0025
Appears in Collections:Obstetrics and Gynaecology publications

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