Meiotic abnormalities in in vitro-matured marmoset monkey (Callithrix jacchus) oocytes: development of a non-human primate model to investigate causal factors.

BACKGROUND: Meiotic abnormalities are thought to be a major causal factor of low embryo development rates, for embryos developed from in vitro-matured oocytes. A new non-human primate model, in the common marmoset, is being developed to facilitate investigation of the mechanisms involved. METHODS: Oocytes were dissected from antral follicles from three size classes. They were allowed to mature in vitro for only 24 h, in order to focus the investigation on the rapidly maturing oocytes. Chromosome spreads were visualized with Giemsa staining, and spindles /chromosomes with fluorescently labelled anti-alpha-tubulin antibody combined with a DNA fluorochrome. RESULTS: 40% of the oocytes had reached metaphase II (MII) after 24 h. Of the MII oocytes selected for karyotyping, readable chromosomal spreads were obtained from 64%. Overall, 63% of these presented a normal haploid chromosome number of 23,X, with all abnormal karyotypes occurring in the oocytes from small follicles. For another group of MII oocytes, where meiotic spindles were visualized, only half of the MII oocytes displayed well-formed spindles and apparently correct chromosomal alignment. CONCLUSIONS: This work provides the first information on the normal and aneuploid MII meiotic chromosome sets for the marmoset oocyte, and demonstrates a high rate of chromosomal and spindle abnormality among rapidly maturing oocytes from small antral follicles.

Comparison of the effects of Percoll and PureSperm on the common marmoset (Callithrix jacchus) semen.

Density gradient centrifugation is a widely used technique for the separation of motile from non-motile sperm, for the removal of contaminating agents such as bacteria and viruses, and for the removal of seminal plasma or cryoprotectant. In the choice of a density gradient medium for a new species, it is important to perform toxicity tests. The present study was carried out to evaluate the potential toxic effects of two silica-based density gradient products (Percoll and PureSperm), on the sperm of the common marmoset. We assessed two different batches of Percoll (polyvinylpyrrolidone (PVP)-coated colloidal silica particles) and one of PureSperm (saline-coated colloidal silica suspension) by means of a computer-aided sperm analysis on semen collected by vibrostimulation. The results showed that although some of the sperm patterns of movement and viability changed significantly over time, and provide a first description of marmoset sperm motility changes under capacitating conditions, there was no significant difference in the sperm treated with Percoll or PureSperm in comparison with the control. We conclude that simple exposure to either of these products does not have a negative effect on viability or motility of marmoset sperm.

Effect of mucin alone and in combination with various dentifrices on in vitro Remineralization.

The aim of this in vitro study was to evaluate the effect of combining various fluoridated dentifrices with mucin on remineralization of bovine enamel. Enamel specimens were embedded in epoxy resin, partly covered with nail varnish, and demineralized in a lactic acid solution (pH 5.0, 14 days). Parts of the demineralized areas of the specimens were then covered with nail varnish. Half of the samples were exposed to a mucin-containing (2.7 g/l) remineralizing solution, the other half to a mucin-free remineralizing solution for 30 days. In each group, the specimens were divided into four subgroups, which were brushed twice a day with a toothpaste containing sodium, stannous/amine, or amine fluoride. The specimens of the fourth subgroup were not brushed, but stored in one of the two solutions. Mineral loss and lesion depth were evaluated from microradiographs. After the remineralization period, specimens that were brushed with one of the dentifrices and stored in the mucin-containing remineralizing solution reacquired more mineral than those brushed and stored in the mucin-free solution (p < 0.05; Bonferroni post hoc test). The results indicate that mucin in combination with various fluorides seems to affect enamel remineralization significantly. Thus, mucin could be considered as an additive to saliva substitutes or mouthwashes in patients with hyposalivation.