Document Detail

Generation of chimeras by morula aggregation.
MedLine Citation:
PMID:  19504074     Owner:  NLM     Status:  MEDLINE    
This chapter describes the tools and the experimental route of targeted manipulation by aggregation in the mouse using targeted embryonic stem cells (ES cells). Instead of injecting ES cells into the blastocoel of a diploid blastocyst-stage embryo (3.5 dpc) ES cells can be brought together with diploid morula-stage embryos (2.5 dpc). The zona pellucida of the embryo needs to be removed and one or two embryos (sandwich aggregation) are put together with ES cells into an indentation well of a cell culture grade dish overnight for aggregation. This can be performed manually using a stereomicroscope and does not require any special training or expensive instrumentation.The next day, the embryo would have developed into a blastocyst in vitro and can be transferred to a pseudopregnant female mouse (see Chapter 15 ).The use of tetraploid embryos generated by electrofusion will lead to entirely ES cell-derived fetuses.
Anne Plück; Christian Klasen
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  561     ISSN:  1064-3745     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2009  
Date Detail:
Created Date:  2009-06-08     Completed Date:  2009-09-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  219-29     Citation Subset:  IM    
Centre for Mouse Genetics, Institute for Genetics, University of Cologne, Koeln, Germany.
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MeSH Terms
Animals, Genetically Modified
Blastocyst / cytology,  physiology
Cell Aggregation / genetics*
Cell Differentiation
Chimera / genetics*
Embryo Culture Techniques
Embryo, Mammalian / cytology*
Embryonic Stem Cells / cytology*,  physiology
Fetal Viability
Microinjections / methods
Morula / cytology*,  physiology
Polymerase Chain Reaction

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