Document Detail


Cell donor influences success of producing cattle by somatic cell nuclear transfer.
MedLine Citation:
PMID:  14998911     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
A M Powell; N C Talbot; K D Wells; D E Kerr; V G Pursel; R J Wall
Publication Detail:
Type:  Journal Article     Date:  2004-03-03
Journal Detail:
Title:  Biology of reproduction     Volume:  71     ISSN:  0006-3363     ISO Abbreviation:  Biol. Reprod.     Publication Date:  2004 Jul 
Date Detail:
Created Date:  2004-06-22     Completed Date:  2005-01-05     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  0207224     Medline TA:  Biol Reprod     Country:  United States    
Other Details:
Languages:  eng     Pagination:  210-6     Citation Subset:  IM    
Affiliation:
Biotechnology and Germplasm Laboratory, Agricultural Research Service, USDA, Beltsville, Maryland 20705, USA.
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MeSH Terms
Descriptor/Qualifier:
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:
0/Culture Media; 7782-44-7/Oxygen

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