Characterization, Cryopreservation, and Ablation of Spermatogonial Stem Cells in Adult Rhesus Macaques

Authors

  • Brian P. Hermann,

    1. Departments of Ob/Gyn & Reproductive Sciences, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA
    2. Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA
    3. Center for Research in Reproductive Physiology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA
    4. Pittsburgh Development Center, Magee-Womens Research Institute & Foundation, Pittsburgh, Pennsylvania, USA
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  • Meena Sukhwani,

    1. Pittsburgh Development Center, Magee-Womens Research Institute & Foundation, Pittsburgh, Pennsylvania, USA
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  • Chih-Cheng Lin,

    1. Pittsburgh Development Center, Magee-Womens Research Institute & Foundation, Pittsburgh, Pennsylvania, USA
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  • Yi Sheng,

    1. Pittsburgh Development Center, Magee-Womens Research Institute & Foundation, Pittsburgh, Pennsylvania, USA
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  • Jamie Tomko,

    1. Pittsburgh Development Center, Magee-Womens Research Institute & Foundation, Pittsburgh, Pennsylvania, USA
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  • Mario Rodriguez,

    1. Pittsburgh Development Center, Magee-Womens Research Institute & Foundation, Pittsburgh, Pennsylvania, USA
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  • Jennifer J. Shuttleworth,

    1. Pittsburgh Development Center, Magee-Womens Research Institute & Foundation, Pittsburgh, Pennsylvania, USA
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  • David McFarland,

    1. Pittsburgh Development Center, Magee-Womens Research Institute & Foundation, Pittsburgh, Pennsylvania, USA
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  • Robin M. Hobbs,

    1. Cancer Biology and Genetics Program and Department of Pathology, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, New York, New York, USA
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  • Pier Paolo Pandolfi,

    1. Cancer Biology and Genetics Program and Department of Pathology, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, New York, New York, USA
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  • Gerald P. Schatten,

    1. Departments of Ob/Gyn & Reproductive Sciences, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA
    2. Cell Biology and Physiology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA
    3. Pittsburgh Development Center, Magee-Womens Research Institute & Foundation, Pittsburgh, Pennsylvania, USA
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  • Kyle E. Orwig Ph.D.

    Corresponding author
    1. Departments of Ob/Gyn & Reproductive Sciences, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA
    2. Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA
    3. Center for Research in Reproductive Physiology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA
    4. Pittsburgh Development Center, Magee-Womens Research Institute & Foundation, Pittsburgh, Pennsylvania, USA
    • University of Pittsburgh School of Medicine, Magee-Womens Research Institute & Foundation, 204 Craft Avenue, Pittsburgh, Pennsylvania 15213, USA. Telephone: 412-641-2460; Fax: 412-641-3899
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Abstract

Spermatogonial stem cells (SSCs) are at the foundation of mammalian spermatogenesis. Whereas rare Asingle spermatogonia comprise the rodent SSC pool, primate spermatogenesis arises from more abundant Adark and Apale spermatogonia, and the identity of the stem cell is subject to debate. The fundamental differences between these models highlight the need to investigate the biology of primate SSCs, which have greater relevance to human physiology. The alkylating chemotherapeutic agent, busulfan, ablates spermatogenesis in rodents and causes infertility in humans. We treated adult rhesus macaques with busulfan to gain insights about its effects on SSCs and spermatogenesis. Busulfan treatment caused acute declines in testis volume and sperm counts, indicating a disruption of spermatogenesis. One year following high-dose busulfan treatment, sperm counts remained undetectable, and testes were depleted of germ cells. Similar to rodents, rhesus spermatogonia expressed markers of germ cells (VASA, DAZL) and stem/progenitor spermatogonia (PLZF and GFRα1), and cells expressing these markers were depleted following high-dose busulfan treatment. Furthermore, fresh or cryopreserved germ cells from normal rhesus testes produced colonies of spermatogonia, which persisted as chains on the basement membrane of mouse seminiferous tubules in the primate to nude mouse xenotransplant assay. In contrast, testis cells from animals that received high-dose busulfan produced no colonies. These studies provide basic information about rhesus SSC activity and the impact of busulfan on the stem cell pool. In addition, the germ cell-depleted testis model will enable autologous/homologous transplantation to study stem cell/niche interactions in nonhuman primate testes.

Disclosure of potential conflicts of interest is found at the end of this article.

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