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Mary Zelinski Laboratory

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Organisms and Viruses

Protocols

  • In vivo contraceptive trials protocol ( Protocol )

    In vivo contraceptive trials – female

    The Nonhuman Primate Contraceptive Core (NPCC) provides services and coordination of animal usage for U54 Contraceptive Center investigators with approved projects. The NPCC maintains a pool of cynomolgus macaque females and males in the Division of Animal Resources (DAR), ONPR, available for contraceptive (mating) trials; coordinates and performs studies on potential contraceptive agents; confirms menstruation and ensures mating has occurred by taking vaginal swabs; performs ultrasounds to monitor for pregnancies; and obtains blood samples for hormone assays, pharmacokinetic studies and metabolic profiles. The Core acclimates the females to a small group (harem) housing situation (8-10 females to one male) wherein they can be separated for daily vaginal checks, dosing, blood sampling and ultrasound analyses.

    The NPCC is uniquely poised to drive proof-of-concept studies in nonhuman primates that can be readily translated into further efforts to test novel contraceptives in women. The final stage in the strategy to discover novel agents for female fertility regulation will be to evaluate the efficacy of these agents as reversible contraceptive agents in a nonhuman primate model. Cost-effective, experimental contraceptive trials can be performed in the NPCC to (1) demonstrate a contraceptive effect of chronic (6 month) administration of the agent in a breeding colony of macaques exhibiting natural menstrual cycles; and (2) document that the contraceptive effect is reversible following discontinuation for a 6 month interval. Data from these translational experiments conducted in the NPCC will increase confidence in the feasibility of considering a continual regimen of these inhibitors for clinical use as a contraceptive in women.

    In vivo contraceptive trials – male

    The Nonhuman Primate Contraceptive Core (NPCC) provides services and coordination of animal usage for U54 Contraceptive Center investigators with approved projects. Male rhesus monkeys are leased from the Oregon National Primate Research Center and trained for semen collection. Proof-of-concept studies based on semen parameters (count, motility, live-dead staining, progressive motion), testicular biopsies, hormone levels and pharmacokinetics can be performed. Contraceptive (mating) trials are coordinated and performed studies using small harem groups of cynomolgus or rhesus female. The NPCC confirms menstruation and ensures mating has occurred by taking vaginal swabs; performs ultrasounds to monitor for pregnancies; and obtains blood samples for hormone assays, pharmacokinetic studies and metabolic profiles. The Core acclimates the females to a small group (harem) housing situation (8-10 females to one male) wherein they can be separated for daily vaginal checks, dosing, blood sampling and ultrasound analyses.

    The NPCC is uniquely poised to drive proof-of-concept studies in nonhuman primates that can be readily translated into further efforts to test novel contraceptives in men. The final stage in the strategy to discover novel agents for male fertility regulation will be to evaluate the efficacy of these agents as reversible contraceptive agents in a nonhuman primate model. Cost-effective, experimental contraceptive trials can be performed in the NPCC to (1) demonstrate a contraceptive effect of chronic (6 month) administration of the agent to males housed in a breeding colony of female macaques exhibiting natural menstrual cycles; and (2) document that the contraceptive effect is reversible following discontinuation for a 6 month interval. Data from these translational experiments conducted in the NPCC will increase confidence in the feasibility of considering a these agents for clinical use as a contraceptive in men.

  • Nonhuman primate contraceptive model protocol ( Protocol )

    Animals were treated in accord with the NIH Guide for the Care and Use of Laboratory Animals and under research protocols reviewed and approved by the institutional animal care and use committee of the ONPRC.
    Anesthetized animals underwent a pre-screening laparoscopy to evaluate ovarian ligaments length. Only those with long ligaments, which were necessary to undergo ovarian manipulations with minimal damage to ovarian circulation, were included in the experiments.
    Each animal was placed horizontally on a stainless steel table, with a hydraulic lift to facilitate positioning. Stationary laser beams were utilized to position the ovaries at a reproducible point in space associated with an exact calibrated position. Ovaries were positioned carefully outside the ventral surface either with a hemostat attached to the ovarian ligament and surrounded by an additional lead collar to protect the rest of the body, or were placed within a shield that tightly collimated the radiation beam down to 13 mm diameter (the ovary was positioned within a small lead cup at the end of the collimator). Ovaries were covered with sterile gauze sponges soaked in warm sterile saline. Irradiation commenced for a pre-determined duration of 13 min to generate 1,500 rads of absorbed dose (see below). The female was then removed from the irradiation table, sutured and returned to her cage. Animals assigned to sham-irradiation underwent surgical manipulations and procedures (i.e., externalization of the ovary for 20 min) without OXI.

  • Oncofertility protocol ( Protocol )

    Publication: PMID: 15014485. Fertility preservation. Lab has ovarian radiation protocol.

  • Ovarian X-irradiation (OXI) description protocol ( Protocol )

    The X-ray machine utilized was a Phillips 324 kVp, and the beam selected for use employed the ISO HK250 technique, which involves an energy spectrum of 122 keV average and 250 keV maximum. The voltage and current settings used were 250 kV and 12 mA, respectively. The beam filtration that provided this spectrum is made up of 1.60 mm copper, including the 4-mm inherent filtration of the machine. The resulting half-value and quarter- value layers were 2.463 mm and 3.37 mm of copper, respectively. This beam was selected based on its relatively high energy combined with high dose rate, which allows good dose uniformity throughout the full depth of the ovary and a quick (13-min total) exposure time. The additional collimator used for the irradiations provided a tight 13 mm diameter beam for the ovary that was within the lead shield device at a distance of 34 cm (distance from the X-ray machine target). The exposure rate and air-kerma rate measured for this beam are traceable to national standards (NIST).

Services

  • Fertility preservation and contraception consultation service ( Support service )

    Research consultation in the following areas:

    - Controlled ovarian stimulation with recombinant human gonadotropins in nonhuman primates
    - Culture and maintenence of preimplantation embryos
    - Cryopreservation of ovarian cortex using slow-freeze or vitrification methods
    - Immunocytochemical analyses in nonhuman primate ovaries
    - In vitro culture of individual macaque preantral follicles isolated from ovarian cortex
    - In vitro fertilization
    - In vitro maturation of cumulus-oocyte complexes
    - Lutenizing granulosa cell purification and culture
    - Monitoring menstrual/ovarian cyclicity in nonhuman primates
    - Morphometric follicle counts in macaque ovaries
    - Oocyte isolation from follicular aspirates and identification of nuclear maturity
    - Semen characterization
    - Semen collection and preparation
    - Ultrasound ovaries, nonpregnant and pregnant uteri, fetal measurements


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Last updated: 2011-04-12T13:25:29.666-05:00

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The eagle-i Consortium is supported by NIH Grant #5U24RR029825-02 / Copyright 2016