| Generation of chimeras by morula aggregation. | |
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MedLine Citation:
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PMID: 19504074 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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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. |
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Authors:
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Anne Plück; Christian Klasen |
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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: 561 ISSN: 1064-3745 ISO Abbreviation: Methods Mol. Biol. Publication Date: 2009 |
Date Detail:
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Created Date: 2009-06-08 Completed Date: 2009-09-22 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: 219-29 Citation Subset: IM |
Affiliation:
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Centre for Mouse Genetics, Institute for Genetics, University of Cologne, Koeln, Germany. Anne.Plueck@uni-koeln.de |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals 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 Mice Microinjections / methods Morula / cytology*, physiology Ploidies Polymerase Chain Reaction |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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