| Cell donor influences success of producing cattle by somatic cell nuclear transfer. | |
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MedLine Citation:
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PMID: 14998911 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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To assess sources of variation in nuclear transfer efficiency, bovine fetal fibroblasts (BFF), harvested from six Jersey fetuses, were cultured under various conditions. After transfection, frozen-thawed lung or muscle BFF donor cells were initially cultured in DMEM in 5% CO(2) and air and some were transferred to MEM, with 5% or 20% O(2) or 0.5% or 10% serum and G418 for 2-3 wk. Selected clonal transfected fibroblasts were fused to enucleated oocytes. Fused couplets (n = 4007), activated with ionomycin and 6-dimethylaminopurine, yielded 927 blastocysts, and 650 were transferred to 330 recipients. Fusion rate was influenced by oxygen tension in a fetus-dependent manner (P < 0.001). Blastocyst development was influenced in a number of ways. Hip fibroblast generated more blastocysts when cultured in MEM (P < 0.001). The influence of serum concentration was fetus dependent (P < 0.001) and exposing fibroblast to low oxygen was detrimental to blastocyst development (P < 0.001). Cells from two of the six fetuses produced embryos that maintained pregnancies to term, resulting in eight viable calves. Pregnancy rates 56 days after transfer for the two productive donor fetuses, was at least double that of other recipients and may provide a fitness indicator of BFF cell sources for nuclear transfer. We conclude that a significant component in determining somatic cell nuclear transfer success is the source of the nuclear donor cells. |
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Authors:
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A M Powell; N C Talbot; K D Wells; D E Kerr; V G Pursel; R J Wall |
Publication Detail:
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Type: Journal Article Date: 2004-03-03 |
Journal Detail:
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Title: Biology of reproduction Volume: 71 ISSN: 0006-3363 ISO Abbreviation: Biol. Reprod. Publication Date: 2004 Jul |
Date Detail:
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Created Date: 2004-06-22 Completed Date: 2005-01-05 Revised Date: 2007-11-15 |
Medline Journal Info:
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Nlm Unique ID: 0207224 Medline TA: Biol Reprod Country: United States |
Other Details:
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Languages: eng Pagination: 210-6 Citation Subset: IM |
Affiliation:
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Biotechnology and Germplasm Laboratory, Agricultural Research Service, USDA, Beltsville, Maryland 20705, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Animals, Newborn Cattle* / embryology Cell Fusion Cloning, Organism / methods* Culture Media Embryo Transfer Embryo, Mammalian / cytology Female Fibroblasts / metabolism Lung / embryology Muscle, Skeletal / embryology Nuclear Transfer Techniques* Oocytes Oxygen / metabolism Pregnancy Pregnancy Rate Tissue Donors* |
| Chemical | |
Reg. No./Substance:
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0/Culture Media; 7782-44-7/Oxygen |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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