| Production of cloned calves using roscovitine-treated adult somatic cells as donors. | |
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MedLine Citation:
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PMID: 16988376 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Kazuchika Miyoshi; Sezen Arat; Steven L Stice |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Methods in molecular biology (Clifton, N.J.) Volume: 348 ISSN: 1064-3745 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2006 |
Date Detail:
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Created Date: 2006-09-21 Completed Date: 2006-12-07 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9214969 Medline TA: Methods Mol Biol Country: United States |
Other Details:
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Languages: eng Pagination: 125-34 Citation Subset: IM |
Affiliation:
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Faculty of Agriculture, Kagoshima, University, Kagoshima, Japan. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cattle* Cells, Cultured Cloning, Organism / methods* Embryo Transfer / veterinary* Female Fertilization in Vitro / veterinary* Granulosa Cells / drug effects Nuclear Transfer Techniques / veterinary Oocytes / physiology Purines / pharmacology* Tissue Donors |
| Chemical | |
Reg. No./Substance:
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0/Purines; 0/roscovitine |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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