316 EVALUATION OF A HORMONAL PROTOCOL FOR LAPAROSCOPIC OVUM PICK-UP IN PREPUBERTAL EWES

2013 ◽  
Vol 25 (1) ◽  
pp. 305
Author(s):  
P. P. M. Teixeira ◽  
L. C. Padilha ◽  
A. S. L. da Silva ◽  
F. F. P. C. Barros ◽  
L. N. Coutinho ◽  
...  
Keyword(s):  
Ovarian Stimulation ◽  
Ovulation Induction ◽  
High Performance ◽  
In Vitro Maturation ◽  
Prostaglandin F2α ◽  
Hormonal Treatment ◽  
Ovarian Response ◽  
The Other ◽  
Genetic Value

In vitro embryo production is a cutting-edge technology in constant evolution and has been used routinely. With the objective of improving genetic value, recent studies have been focused on the production of high-performance descendants from early prepubertal animals. The aim of this study was to compare hormonal protocols of ovarian stimulation for laparoscopic ovum pick-up in prepubertal sheep, to determine ovarian response and number of oocytes. For this study, 36 Santa Ines sheep, aged between 4 and 8 weeks, were submitted to a progestogen-based, short-term ovulation induction protocol (without use of prostaglandin F2α), and following 36 h, ovarian stimulation (300 IU of eCG) associated with different regimens of administration of FSH, according to experimental group. The ewes were divided into 1 of 6 groups: 2 groups received 80 mg of FSH by either single administration in the first group (G80U) or multiple constant administrations at 12-h intervals in the second group (G80M); animals belonging to 2 other groups received 160 mg in the same fashion, in a single-dose regimen (group G160U) or in multiple administrations (group G160M). The other 2 groups constituted the control groups: one received no hormonal treatment for ovulation induction and ovarian stimulation (GCN); the other received the ovulation induction protocol, but no ovulation stimulation was carried out (GCI). The animals were submitted to laparoscopic ovum pick-up 48 h following the beginning of the ovarian stimulation. The number of follicles viewed (FV), aspirated (FA), and recovered (OR) laparoscopically was recorded. Data were assessed using the one-way ANOVA test, and the comparison among groups was carried out using the Tukey test. The mean of follicles visualised, aspirated, and oocytes recovered are shown in Table 1. Group G160U had a better result regarding OR compared with the other groups. Moreover, the current study highlights that laparoscopic ovum pick-up for in vitro maturation using prepubertal ewes is viable for commercial purposes. More studies are required in order to improve the quality of oocytes and successful in vitro maturation rates. Table 1.Results of visualised follicles (VF), aspirated follicles (AF), and oocytes retrieved (OR)

331 EFFECT OF SUCESSIVE LAPAROSCOPIC OOCYTE RECOVERY AFTER HORMONAL TREATMENT IN CROSSBRED GOATS

2010 ◽  
Vol 22 (1) ◽  
pp. 321
Author(s):  
S. R. G. Avelar ◽  
K. C. Almeida ◽  
A. F. Pereira ◽  
F. C. Sousa ◽  
R. R. Moura ◽  
...  
Keyword(s):  
Ovarian Stimulation ◽  
Prostaglandin F2α ◽  
Hormonal Treatment ◽  
Ovarian Response ◽  
Cumulus Cells ◽  
Embryo Production ◽  
Hormonal Stimulation ◽  
Exact Test ◽  
Oocyte Recovery

Laparoscopic oocyte recovery (LOR) is a valuable tool for obtaining oocytes for in vitro embryo production. When preceded by a treatment of ovarian stimulation, this technique produces an increase in the amount of oocytes recovered. However, a little information has been found to respect the effect of successive hormonal treatments on both oocyte quantity and quality. Therefore, the objectives of this study were to evaluate the ovarian response and quantitative and qualitative COC production. Five adult crossbred goats were hormonally treated with intravaginal sponges containing 60 mg of medroxyprogesterone acetate (MAP, Progespon, Syntex, Buenos Aires, Argentina) for 11 days. In the 8th day of progestagen treatment, 50 μg of prostaglandin F2α analogue (Ciosin, Coopers, São Paulo, Brazil) was administered by i.m. injection. At this time, ovarian stimulation was initiated by the administration of 120 mg pFSH (Folltropin-V, Vetrepharm, Canada) distributed in five decreasing doses (30/30, 20/20, 20 mg), at 12-h intervals. The animals were fasted for 24 h before the laparoscopic procedure, which was performed just after the sponge removal. A laparoscopic system connected to a 22-gauge needle (WTA, Watanabe, Brazil) and a vacuum pump (Biovacuum, Biocom, Brazil) providing 30 mm Hg was used. All follicles with a size larger than 2 mm present in both ovaries were counted and aspirated. The collection medium was TCM-199 supplemented with HEPES (10 mM), heparin (20 IU mL-1), and gentamicin (40 μg mL-1). The COCs were graded based on presence of cumulus cells and cytoplasm homogeneity (I to IV). The hormonal treatment and LOR procedures were repeated three times at 14-day intervals. Data were expressed in percentage or mean ± SEM. The differences were analyzed using ANOVA and Tukey’s or Fischer’s exact test when appropriate, with P < 0.05. No statistical differences were found (P > 0.05) for the number of follicles obtained in each LOR session: 17.0 ± 3.91, 18.75 ± 2.59, and 18.0 ± 4.73, respectively. Repeated LOR procedures also did not affect (P > 0.05) the number of aspirated follicle (15.0 ± 3.92, 15.5 ± 2.33, and 16.0 ± 4.36), resulting from the three sessions, respectively. Average recovery rates were not statistically different (P > 0.05), resulting in 74.7%, 62.9%, and 64.6% between sessions. With respect to the percentage of viable COCs (GI and GII) were not observed statistical differences (P > 0.05), as verified the follow values at 1st to 3rd sessions: 76.79%, 84.62%, and 74.19%. In conclusion, three successive hormonal stimulation LOR procedures did not cause detrimental effects on quantitative and qualitative oocyte production, suggesting that this protocol can be used for programs of in vitro goat embryo production. This study was supported the following Brazilian agencies: FINEP, CNPq, FUNCAP, and CAPES.

scholarly journals Ex vivo Retrieval of Mature Oocytes for Fertility Preservation in a Patient with Bilateral Borderline Ovarian Tumor

2021 ◽  
Author(s):  
Bruno Ramalho de Carvalho ◽  
Geórgia Fontes Cintra ◽  
Taise Moura Franceschi ◽  
Íris de Oliveira Cabral ◽  
Leandro Santos de Araújo Resende ◽  
...  
Keyword(s):  
Fertility Preservation ◽  
Ovarian Stimulation ◽  
Ovarian Tumor ◽  
Ex Vivo ◽  
Oocyte Retrieval ◽  
Ovarian Response ◽  
Malignant Cell ◽  
Borderline Ovarian Tumor ◽  
Increased Risk

AbstractWe report a case of ultrasound-guided ex vivo oocyte retrieval for fertility preservation in a woman with bilateral borderline ovarian tumor, for whom conventional transvaginal oocyte retrieval was deemed unsafe because of the increased risk of malignant cell spillage. Ovarian stimulation with gonadotropins was performed. Surgery was scheduled according to the ovarian response to exogenous gonadotropic stimulation; oophorectomized specimens were obtained by laparoscopy, and oocyte retrieval was performed ∼ 37 hours after the ovulatory trigger. The sum of 20 ovarian follicles were aspirated, and 16 oocytes were obtained. We performed vitrification of 12 metaphase II oocytes and 3 oocytes matured in vitro. Our result emphasizes the viability of ex vivo mature oocyte retrieval after controlled ovarian stimulation for those with high risk of malignant dissemination by conventional approach.

Collection and in vitro maturation of Mazama gouazoubira (brown brocket deer) oocytes obtained after ovarian stimulation

Zygote ◽  
2021 ◽  
pp. 1-7
Author(s):  
Luciana Diniz Rola ◽  
Eveline dos Santos Zanetti ◽  
Maite del Collado ◽  
Ellen de Fátima Carvalho Peroni ◽  
José Maurício Barbanti Duarte
Keyword(s):  
Oocyte Maturation ◽  
Ovarian Stimulation ◽  
In Vitro Maturation ◽  
Wildlife Conservation ◽  
Estradiol Benzoate ◽  
Ex Situ ◽  
Follicular Aspiration ◽  
Mazama Gouazoubira

Summary In vitro production of embryos has gained prominence as a tool for use in wildlife conservation programmes in situ and ex situ. However, the development of this technique depends on steps that include ovarian stimulation, collection and oocyte maturation. The purpose of this study was to assess the feasibility of an ovarian stimulation protocol for follicular aspiration, the efficiency of videolaparoscopy for follicular aspiration and test a medium for in vitro oocyte maturation for the species Mazama gouazoubira. Five females were submitted to repeated ovarian stimulation (hormone protocol using controlled internal drug release), and estradiol benzoate on D0 and eight injections of follicle-stimulating hormone, once every 12 h, from D4 onwards at 30-day intervals. Fourteen surgical procedures were performed in superstimulated females, resulting in the collection of 94 oocytes and an average of 17.1 ± 9.1 follicles observed, 13.5 ± 6.6 follicles aspirated and 7.2 ± 3.7 oocytes collected per surgery. After collection, the oocytes were submitted to in vitro maturation for 24 h and stained with Hoechst 33342 dye to evaluate their nuclear status; 64.5% of the oocytes reached MII and 16.1% were spontaneously activated by parthenogenesis. The nuclear status of oocytes that did not undergo in vitro maturation was evaluated; 80.9% were found to be immature.

scholarly journals Discovery and Preclinical Development of Orally Active Small Molecules that Exhibit Highly Selective Follicle Stimulating Hormone Receptor Agonism

2021 ◽  
Vol 11 ◽  
Author(s):  
Selva Nataraja ◽  
Henry Yu ◽  
Joie Guner ◽  
Stephen Palmer
Keyword(s):  
Ovarian Stimulation ◽  
Hormone Receptor ◽  
Ovulation Induction ◽  
Low Dose ◽  
Intrauterine Insemination ◽  
Follicle Stimulating Hormone ◽  
Controlled Ovarian Stimulation ◽  
Orally Active ◽  
Stimulating Hormone

An orally active follicle stimulating hormone receptor allosteric agonist would provide a preferred treatment for over 16 million infertile women of reproductive age in low complexity methods (ovulation induction-intrauterine insemination) or in high complexity methods (controlled ovarian stimulation-in vitro fertilization). We present two oral follicle stimulating hormone receptor allosteric agonist compounds that have the desired pharmacology, drug metabolism, pharmacokinetics, and safety profile for clinical use. These molecules provide a single agent suitable for ovulation induction-intrauterine insemination or controlled ovarian stimulation-in vitro fertilization that is more convenient for patients and achieves similar preclinical efficacy as rec-hFSH. TOP5668, TOP5300 were evaluated in vitro in Chinese hamster ovary cells transfected with individual glycoprotein receptors measuring cAMP (FSHR, LH/CGR, thyroid stimulating hormone receptor). TOP5668 was found to have solely follicle stimulating hormone receptor allosteric agonist activity while TOP5300 was found to have mixed follicle stimulating hormone receptor allosteric agonist and LHR-AA activity. Both compounds stimulated concentration-dependent increases in estradiol production from cultured rat granulosa cells in the presence or absence of low dose rec-hFSH, while only TOP5300 stimulated testosterone production from rat primary Leydig cells. In pooled human granulosa cells obtained from patients undergoing controlled ovarian stimulation-in vitro fertilization, TOP5300 stimulated 7-fold greater maximal estradiol response than rec-hFSH and TOP5668 was 10-fold more potent than TOP5300. Both TOP5300 and TOP5668 stimulated follicular development in immature rat to the same efficacy as recombinant follicle stimulating hormone. In mice treated with TOP5300, in the presence of low dose of follicle stimulating hormone, there were no differences in oocyte number, fertilization rate, and hatched blastocyst rate in mice with TOP5300 and low dose follicle stimulating hormone vs. reference proteins pregnant mare serum gonadotropin or high dose rec-hFSH. ADME/PK and safety profiles were favorable. In addition, there was no appreciable activity on thyroid hormones by TOP5300 in 14-days toxicological study in rat or dog. The selected lead compound, TOP5300 stimulated a more robust increase in estradiol production from granulosa-lutein cells from women with polycystic ovarian syndrome patient compared to rec-hFSH. Conclusions: Two novel oral FSHR allosteric agonist, TOP5668 and TOP5300, were found to mimic the biological activity of rec hFSH in preclinical studies. Both compounds led to folliculogenesis and superovulation in rat and mice. Specifically, TOP5300 led to a similar number of ovulated oocytes that fertilized and developed into hatched blastocysts in mice when compared to rec-hFSH. The safety profile demonstrated lack of toxicity.

328 IN VITRO MATURATION OF OOCYTES FROM Canindé GOATS SUBMITTED TO DIFFERENT HORMONAL TREATMENTS FOR OVARIAN STIMULATION

2010 ◽  
Vol 22 (1) ◽  
pp. 320
Author(s):  
K. C. Almeida ◽  
A. F. Pereira ◽  
A. S. Alcântara Neto ◽  
S. R. G. Avelar ◽  
F. C. Sousa ◽  
...  
Keyword(s):  
Ovarian Stimulation ◽  
Polar Body ◽  
Hormonal Treatment ◽  
Oocyte Quality ◽  
Exact Test ◽  
Gentamicin Sulfate ◽  
First Polar Body ◽  
The One ◽  
Hormonal Treatments

Oocyte IVM is a long process during which oocytes acquire their ability to support the stages of development in a stepwise manner, ultimately reaching activation of the embryonic genome. The overall success of this process can be affected by factors such as hormonal treatment for ovarian stimulation. Thus, the current study aims to evaluate the possible effects of the ovarian stimulatory protocols on the goat oocyte quality and IVM rate. Adult and cyclic Canindé goats were heat-synchronized by means of intravaginal sponges impregnated with 60 mg medroxyprogesterone acetate (MAP, Progespon, Syntex, Buenos Aires, Argentina) inserted for 11 days coupled with a luteolytic injection of 50 μg cloprostenol (Ciosin, Coopers, São Paulo, Brazil) in the 8th day of treatment. The ovarian stimulation was carried out using one of the following protocols: a) standard multi-doses (MD) with 120 mg pFSH (Folltropin-V, Vetrepharm, Canada) distributed in five injections (30/30; 20/20; 20 mg) at 12 h intervals (n = 18); b) three- doses (TD) with 120 mg pFSH administered in three injections (60; 40; 20 mg) at 24 h intervals (n = 17); c) one shot (OD) of 70 mg pFSH plus 200 IU of eCG (Novormon, Syntex) administered 36 h before sponge removal (n = 17). In MD andTD groups, the pFSH injections started in Day 8 of progestagen treatment. The follicles were aspirated just after the sponge removal using laparoscopic oocyte recovery (LOR). This procedure was performed with a 22-gauge needle and a vacuum pump at 30 mmHg. The collection medium was TCM-199 supplemented with HEPES (10 mM), heparin (20 IU mL-1), and gentamicin sulfate (40 μg mL-1). COCs were classified as grade I, II, III, or IV based on visual criteria (Baldassarre H et al. 2003 Theriogenology 56, 831-839). Good quality oocytes (grade I and II) were incubated in TCM-199 supplemented with cysteamine (100 μM), EGF (10 ng mL-1) and gentamicin sulfate (40 μgm L-1) at 38.5°C in a humidified atmosphere with 5% CO2 in air for 24 h. Oocyte maturation was assessed by the visualization of first polar body under inverted microscope. Data were expressed as percentages and analyzed using the Fischer’s exact test. No statistical differences among hormonal treatments (P > 0.05) were observed for the percentage of the good quality oocytes, with 70.4 ± 3.0% of COCs graded in I and II. The IVM rate inTD (31.4%) was statistically lower than MD (31.4% v. 46.5%, P = 0.04) group. However, no significant differences (P = 0.89) were observed between OD (45.2%) and MD groups. Thus, current results indicate that oocyte production for IVM can be facilitated using ovarian stimulation with the one shot FSH/eCG regime without affecting meiotic competence. In summary, OD and MD treatments can be used for oocyte IVM in an embryo production programme in Canindé goats. This study was supported by the following Brazilian agencies: FINEP, CNPq, FUNCAP, and CAPES.

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