Low temperature storage of rhesus monkey spermatozoa and fertility evaluation by intracytoplasmic injection

Richard R. Yeoman, Shoukhrat Mitalipov, Behzad Gerami-Naini, Kevin D. Nusser, Don P. Wolf

    Research output: Contribution to journalArticle

    15 Citations (Scopus)

    Abstract

    The objective was to develop a sperm freezing procedure suitable for use in the propagation of valuable founder animals by assisted reproductive technologies. Here, we report a comparison of processing methods by measuring the motility of fresh and frozen-thawed rhesus monkey spermatozoa and fertility via intracytoplasmic spermatozoa injection (ICSI) of sibling oocytes. Washed spermatozoa were frozen in straws or in pellets using different cryoprotective media and processed post-thaw with or without a density gradient centrifugation step. Among the four study series, motility post-thaw was improved with density gradient centrifugation (17-24% versus 75%, P <0.01) achieving levels similar to fresh spermatozoa. Spermatozoa injected oocytes (total n = 377) were co-cultured on BRL cells and observed for fertilization and development. With spermatozoa frozen in straws in liquid nitrogen vapors, the fertilization rate after ICSI was lower than with fresh spermatozoa (40-44% versus 77-86%, P <0.05), even with the Percoll-enriched fraction that exhibited robust motility. In contrast, somewhat slower freezing of spermatozoa in pellets on dry ice supported fertilization rates (73%) that were similar to the fresh counterpart. Developmental rates of fertilized eggs were similar in all experiments. A total of 106 embryo transfers has resulted in the first primate born after ICSI with F/T ejaculated spermatozoa plus 22 other infants to date. Additionally, a 3-4 h incubation after thawing improved the fertilization rate with spermatozoa from a male with poor post-thaw recovery of sperm motility. In conclusion, an acceptable fertilization rate after ICSI with motile, frozen-thawed primate spermatozoa was observed comparable to that obtained with fresh spermatozoa allowing small quantities of competent spermatozoa to be used with ICSI to facilitate propagation of desirable primate genotypes.

    Original languageEnglish (US)
    Pages (from-to)2356-2371
    Number of pages16
    JournalTheriogenology
    Volume63
    Issue number9
    DOIs
    StatePublished - Jun 2005

    Fingerprint

    Macaca mulatta
    storage temperature
    Fertility
    Spermatozoa
    spermatozoa
    injection
    Injections
    Temperature
    fertilization (reproduction)
    Fertilization
    Primates
    Density Gradient Centrifugation
    Freezing
    Oocytes
    pellets
    straw
    freezing
    oocytes
    Dry Ice
    assisted reproductive technologies

    Keywords

    • Cryopreservation
    • Fertilization
    • ICSI
    • Rhesus monkey
    • Spermatozoa

    ASJC Scopus subject areas

    • Animal Science and Zoology
    • veterinary(all)

    Cite this

    Low temperature storage of rhesus monkey spermatozoa and fertility evaluation by intracytoplasmic injection. / Yeoman, Richard R.; Mitalipov, Shoukhrat; Gerami-Naini, Behzad; Nusser, Kevin D.; Wolf, Don P.

    In: Theriogenology, Vol. 63, No. 9, 06.2005, p. 2356-2371.

    Research output: Contribution to journalArticle

    Yeoman, Richard R. ; Mitalipov, Shoukhrat ; Gerami-Naini, Behzad ; Nusser, Kevin D. ; Wolf, Don P. / Low temperature storage of rhesus monkey spermatozoa and fertility evaluation by intracytoplasmic injection. In: Theriogenology. 2005 ; Vol. 63, No. 9. pp. 2356-2371.
    @article{bd39c5af28c24bcca960d4e278fecc1f,
    title = "Low temperature storage of rhesus monkey spermatozoa and fertility evaluation by intracytoplasmic injection",
    abstract = "The objective was to develop a sperm freezing procedure suitable for use in the propagation of valuable founder animals by assisted reproductive technologies. Here, we report a comparison of processing methods by measuring the motility of fresh and frozen-thawed rhesus monkey spermatozoa and fertility via intracytoplasmic spermatozoa injection (ICSI) of sibling oocytes. Washed spermatozoa were frozen in straws or in pellets using different cryoprotective media and processed post-thaw with or without a density gradient centrifugation step. Among the four study series, motility post-thaw was improved with density gradient centrifugation (17-24{\%} versus 75{\%}, P <0.01) achieving levels similar to fresh spermatozoa. Spermatozoa injected oocytes (total n = 377) were co-cultured on BRL cells and observed for fertilization and development. With spermatozoa frozen in straws in liquid nitrogen vapors, the fertilization rate after ICSI was lower than with fresh spermatozoa (40-44{\%} versus 77-86{\%}, P <0.05), even with the Percoll-enriched fraction that exhibited robust motility. In contrast, somewhat slower freezing of spermatozoa in pellets on dry ice supported fertilization rates (73{\%}) that were similar to the fresh counterpart. Developmental rates of fertilized eggs were similar in all experiments. A total of 106 embryo transfers has resulted in the first primate born after ICSI with F/T ejaculated spermatozoa plus 22 other infants to date. Additionally, a 3-4 h incubation after thawing improved the fertilization rate with spermatozoa from a male with poor post-thaw recovery of sperm motility. In conclusion, an acceptable fertilization rate after ICSI with motile, frozen-thawed primate spermatozoa was observed comparable to that obtained with fresh spermatozoa allowing small quantities of competent spermatozoa to be used with ICSI to facilitate propagation of desirable primate genotypes.",
    keywords = "Cryopreservation, Fertilization, ICSI, Rhesus monkey, Spermatozoa",
    author = "Yeoman, {Richard R.} and Shoukhrat Mitalipov and Behzad Gerami-Naini and Nusser, {Kevin D.} and Wolf, {Don P.}",
    year = "2005",
    month = "6",
    doi = "10.1016/j.theriogenology.2004.05.033",
    language = "English (US)",
    volume = "63",
    pages = "2356--2371",
    journal = "Theriogenology",
    issn = "0093-691X",
    publisher = "Elsevier Inc.",
    number = "9",

    }

    TY - JOUR

    T1 - Low temperature storage of rhesus monkey spermatozoa and fertility evaluation by intracytoplasmic injection

    AU - Yeoman, Richard R.

    AU - Mitalipov, Shoukhrat

    AU - Gerami-Naini, Behzad

    AU - Nusser, Kevin D.

    AU - Wolf, Don P.

    PY - 2005/6

    Y1 - 2005/6

    N2 - The objective was to develop a sperm freezing procedure suitable for use in the propagation of valuable founder animals by assisted reproductive technologies. Here, we report a comparison of processing methods by measuring the motility of fresh and frozen-thawed rhesus monkey spermatozoa and fertility via intracytoplasmic spermatozoa injection (ICSI) of sibling oocytes. Washed spermatozoa were frozen in straws or in pellets using different cryoprotective media and processed post-thaw with or without a density gradient centrifugation step. Among the four study series, motility post-thaw was improved with density gradient centrifugation (17-24% versus 75%, P <0.01) achieving levels similar to fresh spermatozoa. Spermatozoa injected oocytes (total n = 377) were co-cultured on BRL cells and observed for fertilization and development. With spermatozoa frozen in straws in liquid nitrogen vapors, the fertilization rate after ICSI was lower than with fresh spermatozoa (40-44% versus 77-86%, P <0.05), even with the Percoll-enriched fraction that exhibited robust motility. In contrast, somewhat slower freezing of spermatozoa in pellets on dry ice supported fertilization rates (73%) that were similar to the fresh counterpart. Developmental rates of fertilized eggs were similar in all experiments. A total of 106 embryo transfers has resulted in the first primate born after ICSI with F/T ejaculated spermatozoa plus 22 other infants to date. Additionally, a 3-4 h incubation after thawing improved the fertilization rate with spermatozoa from a male with poor post-thaw recovery of sperm motility. In conclusion, an acceptable fertilization rate after ICSI with motile, frozen-thawed primate spermatozoa was observed comparable to that obtained with fresh spermatozoa allowing small quantities of competent spermatozoa to be used with ICSI to facilitate propagation of desirable primate genotypes.

    AB - The objective was to develop a sperm freezing procedure suitable for use in the propagation of valuable founder animals by assisted reproductive technologies. Here, we report a comparison of processing methods by measuring the motility of fresh and frozen-thawed rhesus monkey spermatozoa and fertility via intracytoplasmic spermatozoa injection (ICSI) of sibling oocytes. Washed spermatozoa were frozen in straws or in pellets using different cryoprotective media and processed post-thaw with or without a density gradient centrifugation step. Among the four study series, motility post-thaw was improved with density gradient centrifugation (17-24% versus 75%, P <0.01) achieving levels similar to fresh spermatozoa. Spermatozoa injected oocytes (total n = 377) were co-cultured on BRL cells and observed for fertilization and development. With spermatozoa frozen in straws in liquid nitrogen vapors, the fertilization rate after ICSI was lower than with fresh spermatozoa (40-44% versus 77-86%, P <0.05), even with the Percoll-enriched fraction that exhibited robust motility. In contrast, somewhat slower freezing of spermatozoa in pellets on dry ice supported fertilization rates (73%) that were similar to the fresh counterpart. Developmental rates of fertilized eggs were similar in all experiments. A total of 106 embryo transfers has resulted in the first primate born after ICSI with F/T ejaculated spermatozoa plus 22 other infants to date. Additionally, a 3-4 h incubation after thawing improved the fertilization rate with spermatozoa from a male with poor post-thaw recovery of sperm motility. In conclusion, an acceptable fertilization rate after ICSI with motile, frozen-thawed primate spermatozoa was observed comparable to that obtained with fresh spermatozoa allowing small quantities of competent spermatozoa to be used with ICSI to facilitate propagation of desirable primate genotypes.

    KW - Cryopreservation

    KW - Fertilization

    KW - ICSI

    KW - Rhesus monkey

    KW - Spermatozoa

    UR - http://www.scopus.com/inward/record.url?scp=19544391236&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=19544391236&partnerID=8YFLogxK

    U2 - 10.1016/j.theriogenology.2004.05.033

    DO - 10.1016/j.theriogenology.2004.05.033

    M3 - Article

    VL - 63

    SP - 2356

    EP - 2371

    JO - Theriogenology

    JF - Theriogenology

    SN - 0093-691X

    IS - 9

    ER -