A comparative approach to somatic cell nuclear transfer in the rhesus monkey

Q. Zhou, S. H. Yang, C. H. Ding, X. C. He, Y. H. Xie, T. B. Hildebrandt, S. M. Mitalipov, X. H. Tang, D. P. Wolf, W. Z. Ji

    Research output: Contribution to journalArticlepeer-review

    36 Scopus citations

    Abstract

    Background: Despite the potential utility of primate somatic cell nuclear transfer (SCNT) to biomedical research and to the production of autologous embryonic stem (ES) cells for cell- or tissue-based therapy, a reliable method for SCNT is not yet available. Employing the rhesus monkey as a clinically relevant animal model, we have compared a conventional electrofusion method for SCNT with a one-step micromanipulation (OSM) method. Methods: A prospective, randomized trial was conducted using only oocytes that were mature [metaphase II (MII)] at collection and a fibroblast-like cell line as nuclear donor cells (fetal fibroblasts). The embryos produced were characterized for in vitro developmental potential, cell number, karyotype and expression of nuclear mitotic apparatus (NuMA) and OCT-4. Results: An in vitro blastocyst development rate of 24.4% was achieved with the OSM method, significantly higher than the 12.2% obtained following electrofusion. SCNT-produced embryos expressed normal karyotypes, cell numbers and NuMA and OCT-4 proteins in most cases. SCNT with male nuclear donor cells resulted in the production of male, SCNT blastocysts, eliminating the possibility of a parthenogenetic origin. Of the four fibroblast cell lines tested as nuclear donor cells, two supported the routine production of blastocysts following SCNT. Conclusions: The application of a modified SCNT technique (OSM) followed by embryo culture in hamster embryo culture medium-10 (HECM-10) allows, for the first time, the routine production of SCNT blastocysts, most of which appear normal by immunochemical, cytochemical and in vitro developmental criteria. These embryos will provide a resource for isolating ES cells and for studies of nuclear reprogramming by monkey cytoplasts.

    Original languageEnglish (US)
    Pages (from-to)2564-2571
    Number of pages8
    JournalHuman Reproduction
    Volume21
    Issue number10
    DOIs
    StatePublished - Oct 2006

    Keywords

    • Rhesus monkey
    • Somatic cell nuclei transfer
    • Therapeutic cloning

    ASJC Scopus subject areas

    • Reproductive Medicine
    • Obstetrics and Gynecology

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