Document Detail


Production of mouse chimeras by injection of embryonic stem cells into the perivitelline space of one-cell stage embryos.
MedLine Citation:
PMID:  20143261     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Generation of mouse chimeras is useful for the elucidation of gene function. In the present report, we describe a new technique for the production of chimeras by injection of R1 embryonic stem (ES) cells into the perivitelline space of one-cell stage mouse embryos. One-cell embryos are injected with 2-6 ES cells into the perivitelline space under the zona pellucida without laser-assistance. Our embryo culture experiments reveal that ES cells injected at the one-cell stage embryo start to be incorporated into the blastomeres beginning at the 8-cell stage and form a chimeric blastocyst after 4 days. We have used this approach to successfully produce a high rate of mouse chimeras in two different mouse genetic backgrounds permitting the establishment of germ line transmitters. This method allows for the earlier introduction of ES cells into mouse embryos, and should free up the possibility of using frozen one-cell embryos for this purpose.
Authors:
Yves De Repentigny; Rashmi Kothary
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2010-02-09
Journal Detail:
Title:  Transgenic research     Volume:  19     ISSN:  1573-9368     ISO Abbreviation:  Transgenic Res.     Publication Date:  2010 Dec 
Date Detail:
Created Date:  2010-11-03     Completed Date:  2011-02-24     Revised Date:  2011-04-28    
Medline Journal Info:
Nlm Unique ID:  9209120     Medline TA:  Transgenic Res     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  1137-44     Citation Subset:  IM    
Affiliation:
Ottawa Hospital Research Institute, 501 Smyth Road, Ottawa, ON, K1H 8L6, Canada.
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MeSH Terms
Descriptor/Qualifier:
Animals
Blastocyst / cytology,  metabolism
Cell Line
Cell Transplantation / methods
Cleavage Stage, Ovum / cytology*,  metabolism
Embryonic Stem Cells / transplantation*
Female
Germ Cells / cytology,  metabolism
Luminescent Proteins / genetics,  metabolism
Male
Mice
Mice, Transgenic
Microinjections
Microscopy, Fluorescence
Pregnancy
Recombinant Proteins / genetics,  metabolism
Transplantation Chimera* / genetics
Grant Support
ID/Acronym/Agency:
//Canadian Institutes of Health Research
Chemical
Reg. No./Substance:
0/Luminescent Proteins; 0/Recombinant Proteins

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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