Document Detail

Production of cloned calves using roscovitine-treated adult somatic cells as donors.
MedLine Citation:
PMID:  16988376     Owner:  NLM     Status:  MEDLINE    
The stage of the donor cell cycle is a major factor in the success of cloning. Quiescent cells arrested in the G0/G1 phases of the cell cycle by either serum starvation or growth arrest when cultured cells reach confluence have been used as donors to produce cloned animals. Recently, we have developed a novel and effective method using roscovitine to synchronize adult bovine granulosa cells in the G0/G1 cell cycle stage. The resulting fetal and calf survival after transfer of cloned embryos was enhanced in the roscovitine-treated group compared with serum-starved controls. The methods described in this chapter outline (1) the preparation of donor cells, (2) the preparation of recipient oocytes, and (3) the production of cloned embryos. The first section involves methods for the preparation of donor cell stocks from isolated granulosa cells and the roscovitine treatment of the cells before nuclear transfer. The second section explains procedures of in vitro maturation of recipient oocytes. The last section involves methods for the production of cell-oocyte complexes, the fusion of the complexes, and the activation, in vitro culture, and transfer into recipient females of cloned embryos.
Kazuchika Miyoshi; Sezen Arat; Steven L Stice
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  348     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2006  
Date Detail:
Created Date:  2006-09-21     Completed Date:  2006-12-07     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  125-34     Citation Subset:  IM    
Faculty of Agriculture, Kagoshima, University, Kagoshima, Japan.
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MeSH Terms
Cells, Cultured
Cloning, Organism / methods*
Embryo Transfer / veterinary*
Fertilization in Vitro / veterinary*
Granulosa Cells / drug effects
Nuclear Transfer Techniques / veterinary
Oocytes / physiology
Purines / pharmacology*
Tissue Donors
Reg. No./Substance:
0/Purines; 0/roscovitine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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