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, Shoukhrat Mitalipov, X. H. Tang, D. P. Wolf, W. Z. Ji

    Research output: Contribution to journalArticle

    35 Citations (Scopus)

    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

    Fingerprint

    Macaca mulatta
    Micromanipulation
    Embryo Transfer
    Embryonic Structures
    Spindle Apparatus
    Fibroblasts
    Blastocyst
    Embryonic Stem Cells
    Karyotype
    Nuclear Transfer Techniques
    Cell Count
    Cell Line
    Metaphase
    Cell- and Tissue-Based Therapy
    Cricetinae
    Primates
    Oocytes
    Haplorhini
    Culture Media
    Biomedical Research

    Keywords

    • Rhesus monkey
    • Somatic cell nuclei transfer
    • Therapeutic cloning

    ASJC Scopus subject areas

    • Physiology
    • Developmental Biology
    • Obstetrics and Gynecology
    • Reproductive Medicine

    Cite this

    Zhou, Q., Yang, S. H., Ding, C. H., He, X. C., Xie, Y. H., Hildebrandt, T. B., ... Ji, W. Z. (2006). A comparative approach to somatic cell nuclear transfer in the rhesus monkey. Human Reproduction, 21(10), 2564-2571. https://doi.org/10.1093/humrep/del216

    A comparative approach to somatic cell nuclear transfer in the rhesus monkey. / Zhou, Q.; Yang, S. H.; Ding, C. H.; He, X. C.; Xie, Y. H.; Hildebrandt, T. B.; Mitalipov, Shoukhrat; Tang, X. H.; Wolf, D. P.; Ji, W. Z.

    In: Human Reproduction, Vol. 21, No. 10, 10.2006, p. 2564-2571.

    Research output: Contribution to journalArticle

    Zhou, Q, Yang, SH, Ding, CH, He, XC, Xie, YH, Hildebrandt, TB, Mitalipov, S, Tang, XH, Wolf, DP & Ji, WZ 2006, 'A comparative approach to somatic cell nuclear transfer in the rhesus monkey', Human Reproduction, vol. 21, no. 10, pp. 2564-2571. https://doi.org/10.1093/humrep/del216
    Zhou Q, Yang SH, Ding CH, He XC, Xie YH, Hildebrandt TB et al. A comparative approach to somatic cell nuclear transfer in the rhesus monkey. Human Reproduction. 2006 Oct;21(10):2564-2571. https://doi.org/10.1093/humrep/del216
    Zhou, Q. ; Yang, S. H. ; Ding, C. H. ; He, X. C. ; Xie, Y. H. ; Hildebrandt, T. B. ; Mitalipov, Shoukhrat ; Tang, X. H. ; Wolf, D. P. ; Ji, W. Z. / A comparative approach to somatic cell nuclear transfer in the rhesus monkey. In: Human Reproduction. 2006 ; Vol. 21, No. 10. pp. 2564-2571.
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    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.",
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    AU - Ding, C. H.

    AU - He, X. C.

    AU - Xie, Y. H.

    AU - Hildebrandt, T. B.

    AU - Mitalipov, Shoukhrat

    AU - Tang, X. H.

    AU - Wolf, D. P.

    AU - Ji, W. Z.

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