Document Detail


Chromosome transfer in mature oocytes.
MedLine Citation:
PMID:  20539289     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
In this article, we describe detailed protocols for the isolation and transfer of spindle-chromosomal complexes between mature, metaphase II-arrested oocytes. In brief, the spindle-chromosomal complex is visualized using a polarized microscope and extracted into a membrane-enclosed karyoplast. Chromosomes are then reintroduced into an enucleated recipient egg (cytoplast), derived from another female, by karyoplast-cytoplast membrane fusion. Newly reconstructed oocytes consist of nuclear genetic material from one female and cytoplasmic components, including mitochondria and mitochondrial DNA (mtDNA), from another female. This approach yields developmentally competent oocytes suitable for fertilization and producing embryonic stem cells or healthy offspring. The protocol was initially developed for monkey oocytes but can also be used in other species, including mouse and human oocytes. Potential clinical applications include mitochondrial gene replacement therapy to prevent transmission of mtDNA mutations and treatment of infertility caused by cytoplasmic defects in oocytes. Chromosome transfer between the cohorts of oocytes isolated from two females can be completed within 2 h.
Authors:
Masahito Tachibana; Michelle Sparman; Shoukhrat Mitalipov
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Publication Detail:
Type:  In Vitro; Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2010-05-27
Journal Detail:
Title:  Nature protocols     Volume:  5     ISSN:  1750-2799     ISO Abbreviation:  Nat Protoc     Publication Date:  2010 Jun 
Date Detail:
Created Date:  2010-06-11     Completed Date:  2010-09-20     Revised Date:  2014-09-17    
Medline Journal Info:
Nlm Unique ID:  101284307     Medline TA:  Nat Protoc     Country:  England    
Other Details:
Languages:  eng     Pagination:  1138-47     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Animals
Chromosomes / genetics,  ultrastructure*
Female
Gene Transfer Techniques
Humans
Macaca mulatta
Male
Metaphase / genetics
Mice
Micromanipulation / instrumentation,  methods
Nuclear Transfer Techniques* / instrumentation
Oocytes / cytology,  ultrastructure*
Sperm Injections, Intracytoplasmic
Grant Support
ID/Acronym/Agency:
HD047675/HD/NICHD NIH HHS; HD047721/HD/NICHD NIH HHS; HD057121/HD/NICHD NIH HHS; HD059946/HD/NICHD NIH HHS; HD063276/HD/NICHD NIH HHS; R01 HD057121/HD/NICHD NIH HHS; R01 HD057121-03/HD/NICHD NIH HHS; R01 HD059946/HD/NICHD NIH HHS; R01 HD059946-03/HD/NICHD NIH HHS; R01 HD063276/HD/NICHD NIH HHS; R01 HD063276-02/HD/NICHD NIH HHS; RR0000163/RR/NCRR NIH HHS; U54 HD18185/HD/NICHD NIH HHS
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