Effects of ovarian stimulation, with and without human chorionic gonadotrophin, on oocyte meiotic and developmental competence in the marmoset monkey (Callithrix jacchus)

Research Centre for Reproductive Health, School of Paediatrics and Reproductive Health, The University of Adelaide, Adelaide, SA 5005, Australia.
Theriogenology (Impact Factor: 1.8). 11/2007; 68(6):861-72. DOI: 10.1016/j.theriogenology.2007.07.009
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A reliable ovarian stimulation protocol for marmosets is needed to enhance their use as a model for studying human and non-human primate oocyte biology. In this species, a standard dose of hCG did not effectively induce oocyte maturation in vivo. The objectives of this study were to characterize ovarian response to an FSH priming regimen in marmosets, given without or with a high dose of hCG, and to determine the meiotic and developmental competence of the oocytes isolated. Ovaries were removed from synchronized marmosets treated with FSH alone (50 IU/d for 6 d) or the same FSH treatment combined with a single injection of hCG (500 IU). Cumulus-oocyte complexes (COCs) were isolated from large (>1.5mm) and small (0.7-1.5mm) antral follicles. In vivo-matured oocytes were subsequently activated parthenogenetically or fertilized in vitro. Immature oocytes were subjected to in vitro maturation and then activated parthenogenetically. Treatment with FSH and hCG combined increased the number of expanded COCs from large antral follicles compared with FSH alone (23.5 +/- 9.3 versus 6.4 +/- 2.7, mean +/- S.E.M.). Approximately 90% of oocytes surrounded by expanded cumulus cells at the time of isolation were meiotically mature. A blastocyst formation rate of 47% was achieved following fertilization of in vivo-matured oocytes, whereas parthenogenetic activation failed to induce development to the blastocyst stage. The capacity of oocytes to complete meiosis in vitro and cleave was positively correlated with follicle diameter. A dramatic effect of follicle size on spindle formation was observed in oocytes that failed to complete meiosis in vitro. Using the combined FSH and hCG regimen described in this study, large numbers of in vivo matured marmoset oocytes could be reliably collected in a single cycle, making the marmoset a valuable model for studying oocyte maturation in human and non-human primates.

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Available from: Christopher Grupen, Apr 04, 2014
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    • "Surprisingly, in the oocyte maturation studies, a strong positive effect is seldom reported [11] and some studies report negative effects [10,12–17]. The most common dose used in the literature is 1 mg/mL, which we also used for our previous studies with marmoset oocyte maturation [1] [2] [4] [5] [18], and published concentrations for rhesus and human oocyte IVM vary from 0.1 to 1 mg/mL [10,16,19–21]. In the present study, effects of no E 2 and 10 mg/mL E 2 were also tested, to create a dose-response series, each step 10 times more than the previous. "
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    ABSTRACT: The aim of the present study was to critically evaluate the effect of different concentrations of estradiol (E2) during IVM of common marmoset (Callithrix jacchus) oocytes from antral follicles. The doses tested were 0, 0.1, 1, or 10 μg/mL E2 (referred to as 0 E2, 0.1 E2, 1 E2, and 10 E2 groups). After a preincubation, the concentration of E2 in IVM drops under oil was approximately 20% of the amount added (0.02; 0.2 and 1.9 μg/mL, respectively) because of absorption into the oil. Oocyte progression to metaphase II was significantly higher in the 0.1 E2 group than that in the absence of E2. With progressively higher doses, the maturation rate tended to decrease suggesting an overdose effect. Furthermore, the total first cleavage rate was significantly higher in the 0.1 E2 group than that in the 0 E2 group and decreased progressively with further increases in E2 concentration, with the 10 E2 group showing the same low rate as without E2. The oocytes which failed to cleave, after maturation in 10 E2, showed obvious signs of overdose with the highest rates of degeneration and abnormal spindle form, and an absence of embryo progression. In contrast to these obvious negative effects on the oocyte, 10 E2 was the only group in which a significant increase in radial cumulus expansion was observed. The concentration 0.1 E2, which is 10 times lower than the most commonly used E2 dose, produced the best results in all oocyte factors evaluated. These results represent the first study for a primate species showing a strong positive effect of E2 on oocyte maturation and embryo development, but only at the optimal concentration, and emphasize the critical limits of the optimal concentration range. Copyright © 2015 Elsevier Inc. All rights reserved.
    Theriogenology 01/2015; 83(8). DOI:10.1016/j.theriogenology.2015.01.012 · 1.80 Impact Factor
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    • "Using 1 H-NMR spectroscopy, this study demonstrated that the metabolomic signatures of porcine FF from small and large antral follicles differ markedly and are influenced by the season. As the developmental potential of oocytes recovered from large follicles has been shown to be greater than that of oocytes recovered from small follicles in numerous species (Crozet et al. 1995, Marchal et al. 2002, Iwata et al. 2004, Kauffold et al. 2005, Lequarre et al. 2005, Bagg et al. 2007, Grupen et al. 2007), the differences found may provide insight into the cellular processes involved during the acquisition of oocyte developmental competence. Similarly, the observed differences may provide clues to the possible cause of reduced oocyte quality in sows during the period of seasonal infertility (Bertoldo et al. 2010, 2011b). "
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    ABSTRACT: The microenvironment of the developing follicle is critical to the acquisition of oocyte developmental competence, which is influenced by several factors including follicle size and season. The aim of this study was to characterise the metabolomic signatures of porcine follicular fluid (FF) collected from good and poor follicular environments, using high resolution proton nuclear magnetic resonance ((1)H-NMR) spectroscopy. Sow ovaries were collected at slaughter, 4 days after weaning, in summer and winter. The contents of small (3-4 mm) and large (5-8 mm) diameter follicles were aspirated and pooled separately for each ovary pair. Groups classified as summer-small (n=8), summer-large (n=15), winter-small (n=9) and winter-large (n=15) were analysed by (1)H-NMR spectroscopy. The concentrations of eleven metabolites differed due to follicle size alone (P<0.05), including glucose, lactate, hypoxanthine and five amino acids. The concentrations of all of these metabolites, except for glucose, were lower in large FF compared with small FF. Significant interactions between follicle size and season were found for the concentrations of glutamate, glycine, N-acetyl groups and uridine. Succinate was the only metabolite that differed in concentration due to season alone (P<0.05). The FF levels of progesterone, androstenedione and testosterone were correlated with the concentrations of most of the metabolites examined. The results indicate there is a distinct shift in follicular glucose metabolism as follicles increase in diameter, and suggest that follicular cells may be more vulnerable to oxidative stress during the summer months. Our findings demonstrate the power of (1)H-NMR spectroscopy to expand our understanding of the dynamic and complex microenvironment of the developing follicle.
    Reproduction 06/2013; DOI:10.1530/REP-13-0142 · 3.17 Impact Factor
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    • "The common marmoset has served as a model for human reproduction because of its 28-day ovarian cycle, multiple, non-seasonal ovulations in each cycle, and small size as well as high fecundity in captivity. Recently, refinement of ovarian stimulation to compensate for the relative insensitivity to human gonadotrophins allowed retrieving large numbers of mature and developmentally competent oocytes [6]. Although the marmoset requires unusually high doses of FSH and hCG for superovulation compared with other non-human primates or humans, a recent in vitro study has confirmed equal effectiveness of human recombinant FSH compared to marmoset FSH on the marmoset FSH receptor [7]. "
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    ABSTRACT: Background Cryopreservation and transplantation of ovarian tissue is one option for re-establishing ovarian function, but optimal conditions for graft sustainment and follicular survival are still considered experimental. The present study aims to analyze the effect of FSH treatment on the resting follicle pool in fresh and cryopreserved primate ovarian tissues following xenografting. Methods Ovarian tissues from adult marmosets were grafted freshly or following cryopreservation to ovarectomized nude mice treated with FSH 25 IU twice daily post transplantation or left untreated as controls. Grafts were retrieved 2 or 4 weeks after transplantation to evaluate the number and morphological appearance of follicles. Results Early start of FSH treatment within 1 week following transplantation partly prevents primordial follicle loss in fresh and frozen-thawed tissues, whereas after a 3 weeks time interval this effect is present only in fresh tissues. A similar positive effect of early, but not later FSH treatment on primary follicles is seen in fresh tissues compared to only marginal effects in frozen-thawed tissues. The percentage of morphologically normal follicles is generally increased in FSH treated tissues, whereas the percentage of primary follicles over all primordial and primary follicles is increased by FSH only in freshly-grafted tissues. Conclusions FSH treatment alleviates depletion of the resting follicle pool and promotes normal follicular morphology both in freshly and frozen-thawed grafted tissues. In previously cryopreserved tissues, applying to most of the tissues intended for clinical use in fertility preservation attempts, its positive effect on primordial follicle numbers and potential graft sustainment is dependent on an early start of treatment within one week of transplantation.
    Reproductive Biology and Endocrinology 11/2012; 10(1):98. DOI:10.1186/1477-7827-10-98 · 2.23 Impact Factor
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