Protocol abstract
Nature Protocols 1, - 1145 - 1153 (2006)
Published online: 7 September 2006 | doi:10.1038/nprot.2006.173
Subject Categories: Genetic modification | Model organisms
Production of chimeras by aggregation of embryonic stem cells with diploid or tetraploid mouse embryos
Guy S Eakin1 & Anna-Katerina Hadjantonakis1
Abstract
The production of mouse chimeras is a common step in the establishment of genetically modified animal strains. Chimeras also provide a powerful experimental tool for following cell behavior during both prenatal and postnatal development. This protocol outlines a simple and economical technique for the production of large numbers of mouse chimeras using traditional diploid morula
diploid embryonic stem (ES) cell aggregations. Additional steps are included to describe the procedures necessary to produce specialized tetraploid chimeras using tetraploid moruladiploid ES cell aggregations. This increasingly popular form of chimera produces embryos of nearly complete ES cell derivation that can be used to speed transgenic production or ask developmental questions. Using this protocol, mouse chimeras can be generated and transferred to pseudopregnant surrogate mothers in a 5-d period.
- Developmental Biology Program, Sloan-Kettering Institute, 1275 York Avenue, New York, New York 10021, USA.
Correspondence to: Anna-Katerina Hadjantonakis1 e-mail: hadj@mskcc.org
nature-products
A-Z product listing
- 0.25% (v/v) trypsin/0.2% (w/v) EDTA in PBS(GIBCO)
- 0.3 M mannitol(Sigma)
- 3.5-, 6- and 10-cm tissue culture dishes(Falcon)
- Acid Tyrode's solution(Sigma)
- AffiGel Blue Gelbeads(Bio-Rad)
- Aggregation needles(BLS)
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