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Logo of nihpaAbout Author manuscriptsSubmit a manuscriptHHS Public Access; Author Manuscript; Accepted for publication in peer reviewed journal;
 
Fertil Steril. Author manuscript; available in PMC 2014 January 16.
Published in final edited form as:
PMCID: PMC3893349
NIHMSID: NIHMS367710

Chromosome transfer in mature oocytes

Masahito Tachibana, M.D., Ph.D.,a Michelle Sparman, M.S.,a and Shoukhrat Mitalipov, Ph.D.a,b,c

Abstract

Objective

To demonstrate step-by-step micromanipulation procedures required for transfer of spindle-chromosomal complexes between mature oocytes.

Design

Video presentation of reproductive biology study.

Setting

In vitro fertilization and embryo manipulation laboratory.

Animal(s)

Rhesus (Macaca mulatta) primates.

Intervention(s)

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.

Main Outcome Measure(s)

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.

Result(s)

Newly reconstructed oocytes consist of nuclear genetic material from one female and cytoplasmic components, including mitochondria and mitochondrial DNA from another.

Conclusion(s)

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.

Keywords: Micromanipulation, chromosome transfer, spindle-chromosomal complex, oocytes

VIDEO

Acknowledgments

Supported by institutional funds from Oregon National Primate Research Center and grants from the National Institutes of Health HD057121, HD059946, HD063276, and 8P51OD011092.

Footnotes

Video is available at http://fertstertforum.com/2012974tachibana/.

M.T. has nothing to disclose. M.S. has nothing to disclose. S.M. has nothing to disclose.

Presented at the American Society for Reproductive Medicine annual meeting, Denver, Colorado, October 26, 2010.

SUGGESTED READING

1. Tachibana M, Sparman M, Sritanaudomchai H, Ma H, Clepper L, Woodward J, et al. Mitochondrial gene replacement in primate offspring and embryonic stem cells. Nature. 2009;461:367–72. [PMC free article] [PubMed]
2. Tachibana M, Sparman M, Mitalipov S. Chromosome transfer in mature oocytes. Nature protocols. 2010;5:1138–47. [PMC free article] [PubMed]