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B A Rodrigues

Publications and source records attributed to B A Rodrigues.

6 recordsLinked to original sources

Responses of canine oocytes to in vitro maturation and in vitro fertilization outcome.

The potential benefits of assisted reproduction techniques, such as in vitro maturation (IVM) and in vitro fertilization (IVF) in canids, are linked to the protection and saving of species threatened by extinction due to worldwide habitat destruction and pollution. In both domestic and wild species, these technologies will form the basis for the next leap in reproductive performance by improving fertility rates in valuable middle-aged females, by improving pregnancy rate in infertile or sub-fertile populations and by rescuing biological material to replenish populations of endangered species. In vitro techniques are supposed to answer the reproductive questions of canids, to introduce new methods for contraception and to compete with artificial insemination (AI) as the major or predominant method of embryo production, oocyte- and embryo cryopreservation and cloning. The causes affecting in vitro meiosis of dog oocytes are likely to be diverse. Incomplete understanding of the events associated with oocyte developmental competence are imputed to species reproductive physiology, medium composition and source of ovarian oocyte population used for in vitro maturation. This review addresses some issues on the current state of in vitro maturation and in vitro fertilization of canine oocytes.

Animals↗

The effect of hyaluronan concentrations in hST-supplemented TCM 199 on in vitro nuclear maturation of bitch cumulus-oocyte complexes.

In this study, the effect of hyaluronan (HA) on in vitro nuclear maturation of bitch oocytes was evaluated. Cumulus-oocyte complexes (COCs) were cultured for 48 h at 37 degrees C in a humidified atmosphere of 5% CO2 in air. Oocytes with one or more layers of intact cumulus cells and dark cytoplasm were allocated to the following treatments: (A) TCM 199 supplemented with 25 mM Hepes/L (v/v), with 10% heat inactivated estrous cow serum (ECS), 50 microg/mL gentamycin, 2.2 mg/mL sodium bicarbonate, 22 microg/mL pyruvic acid, 20 microg/mL estradiol, 0.5 microg/mL FSH, 0.03 IU/mL hCG and 1 microg/mL human somatotropin (hST) (control medium), (B) Treatment A + 0.5 mg/mL HA and (C) Treatment A + 1.0 mg/mL HA. Supplementation with HA did not increase the number of oocytes that resumed meiosis. Additionally, there were no differences among treatments in the cumulus cell expansion of oocytes. In conclusion, the addition of HA to hST-supplemented TCM 199 did not improve the in vitro nuclear maturation of bitch oocytes, and therefore appeared to be unsatisfactory as supplement for in vitro maturation (IVM) of canine oocytes.

Animals↗

Meiotic response of in vitro matured canine oocytes under different proteins and heterologous hormone supplementation.

The impact of TCM-199 supplemented with different proteins and heterologous hormones on the in vitro maturation (IVM) rate of bitch oocytes was evaluated by nuclear staining under fluorescence microscopy. Oocytes were recovered by slicing of ovaries from bitches presented at various stages of oestrous cycle to ovariohysterectomy. The basic culture medium was TCM-199 supplemented with 25 mM Hepes/l, with 10% heat-inactivated oestrous cow serum (ECS), 50 microg/ml gentamicin, 2.2 mg/ml sodium bicarbonate and 22-microg/ml pyruvic acid, 1.0-microg/ml oestradiol (E 8875; Sigma), 0.5-microg/ml follicle-stimulating hormone (FSH) (Folltropin-V; Vetrepharm Inc., Ontario, Canada) and 0.03 IU/ml human gonadotropin (hCG) (Profasi HP; Serono, Aubonne, Switzerland). Oocytes were distributed randomly between basic culture medium (control) and the corresponding experimental treatment. Hormone treatments were: oocytes cultured in; (1) medium without FSH, (2) control medium supplemented with 20 microg/ml oestradiol, or (3) medium supplemented with 1 microg/ml human somatotropin (hST; Humatrope, Lilly, Saint Cloud, France). The second experiment consisted of oocytes cultured in medium supplemented with 0.4% (w/v) bovine serum albumin (BSA, fraction V; Gibco Grand Island, NY, USA) instead of ECS, or oocytes cultured in medium with 10% inactivated oestrous bitch serum (EBS) instead of ECS. Oocytes were cultured in 100 microl droplets (up to 25 oocytes per drop) under mineral oil at 37 degrees C in a 100% humidified atmosphere containing 5% CO2 in air. After 72 h of IVM, the highest rates (p < 0.05) of meiotic resumption were achieved with the 0.4% BSA supplementation. A positive influence on the metaphase II (MII) acquisition rate was observed with hST supplement. Oocytes cultured with 10% EBS supplementation did not develop to the MII stage. The results in this study show that the protein and hormone supplements to TCM-199 culture medium tested did not promote the final steps of IVM of bitch oocytes.

Animals↗