Chromosome transfer in mature oocytes.

Masahito Tachibana, Michelle Sparman, Shoukhrat Mitalipov

    Research output: Contribution to journalArticle

    23 Citations (Scopus)

    Abstract

    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.

    Original languageEnglish (US)
    Pages (from-to)1138-1147
    Number of pages10
    JournalNature Protocols
    Volume5
    Issue number6
    DOIs
    StatePublished - 2010

    Fingerprint

    Chromosomes
    Mitochondrial DNA
    Oocytes
    Membranes
    Mitochondria
    Stem cells
    Microscopes
    Fusion reactions
    Genes
    Defects
    Membrane Fusion
    Mitochondrial Genes
    Metaphase
    Embryonic Stem Cells
    Fertilization
    Genetic Therapy
    Infertility
    Haplorhini
    Ovum
    Mutation

    ASJC Scopus subject areas

    • Biochemistry, Genetics and Molecular Biology(all)

    Cite this

    Chromosome transfer in mature oocytes. / Tachibana, Masahito; Sparman, Michelle; Mitalipov, Shoukhrat.

    In: Nature Protocols, Vol. 5, No. 6, 2010, p. 1138-1147.

    Research output: Contribution to journalArticle

    Tachibana, Masahito ; Sparman, Michelle ; Mitalipov, Shoukhrat. / Chromosome transfer in mature oocytes. In: Nature Protocols. 2010 ; Vol. 5, No. 6. pp. 1138-1147.
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