title: Chromosome transfer in mature oocytes.
abstract: OBJECTIVE
  To demonstrate step-by-step micromanipulation procedures required for transfer of spindle-chromosomal complexes between mature oocytes.
  
  
  METHODS
  Video presentation of reproductive biology study.
  
  
  METHODS
  In vitro fertilization and embryo manipulation laboratory.
  
  
  METHODS
  Rhesus (Macaca mulatta) primates.
  
  
  METHODS
  Transplantation of the genetic material between mammalian oocytes offers many opportunities to study various aspects of nuclear-cytoplasmic interactions during oogenesis, fertilization and embryo development. We demonstrate the feasibility of isolation and transfer of chromosomes between mature metaphase II (MII) primate oocyte. After fertilization, manipulated oocytes were capable of producing healthy offspring or embryonic stem cells.
  
  
  METHODS
  In this video, we show micromanipulation procedures required for isolation and transfer of spindle-chromosomal complexes between rhesus MII oocytes. In brief, the spindle is visualized using a polarized microscope and extracted into a membrane enclosed karyoplast. Karyoplasts are then reintroduced into an enucleated recipient oocyte (cytoplast, derived from an another female) by karyoplast-cytoplast membrane fusion.
  
  
  RESULTS
  Newly reconstructed oocytes consist of nuclear genetic material from one female and cytoplasmic components, including mitochondria and mitochondrial DNA from another.
  
  
  CONCLUSIONS
  This video demonstrates the protocol developed for primate oocytes that successfully allowed of isolation and transfer of chromosomes between mature metaphase II (MII) 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. Video is available at http://fertstertforum.com/2012974tachibana/.
authors.affiliation: Division of Reproductive and Developmental Sciences, Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, Oregon, USA.
authors.forename: Masahito
authors.initials: M
authors.lastname: Tachibana
authors.forename: Michelle
authors.initials: M
authors.lastname: Sparman
authors.forename: Shoukhrat
authors.initials: S
authors.lastname: Mitalipov
mesh_terms: D000818:Animals; D002875:Chromosomes / Q000648:ultrastructure*; D005240:Feasibility Studies; D005260:Female; D018014:Gene Transfer Techniques; D008253:Macaca mulatta; D008561:Membrane Fusion; D008677:Metaphase; D008846:Micromanipulation; D008859:Microscopy, Polarization; D009865:Oocytes / Q000648:ultrastructure*; D008941:Spindle Apparatus / Q000637:transplantation* / Q000648:ultrastructure