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PubMed · 11900643

Somatic cell cloning without micromanipulators.

G Vajta·I M Lewis·P Hyttel·G A Thouas·A O Trounson
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Abstract

Until now, micromanipulators have been regarded as indispensable for somatic cell nuclear transfer. This paper describes an improved zona-free nuclear transfer procedure with manual bisection of oocytes, selection of cytoplasts by Hoechst staining, and two-step fusion of somatic cells from primary granulosa cell cultures with two cytoplasts. Blastocyst rates in the three systems tested for zona-free embryo culture were 0%, 18%, and 36% for microdrops, well of the wells (WOW system), and microcapillaries (GO system), respectively. This simple, rapid, and inexpensive procedure may become a useful alternative to the existing techniques for somatic cell nuclear transfer for large-scale application of the technology.

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AnimalsBlastocystCattleCell DivisionCell NucleusCloning, OrganismCulture MediaCytoplasmEmbryo, MammalianTime FactorsJournal ArticleResearch Support, Non-U.S. Gov't

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G Vajta, I M Lewis, P Hyttel, G A Thouas, A O Trounson. 2001. Somatic cell cloning without micromanipulators.. https://doi.org/10.1089/15204550152475590

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