Avaliou-se o efeito do soro de cadela em estro na maturação in vitro de ovócitos caninos, utilizando-se 92 ovócitos de cadelas, submetidas à cirurgia eletiva de ovarioisterectomia. Os ovócitos foram selecionados e distribuídos em dois tratamentos: T1 (n = 48), ovócitos cultivados in vitro durante 96 horas utilizando meio base - TCM199 + 5µg/mL de LH + 20µg/mL de FSH - mais 10% de soro inativado de vaca em estro e T2 (n = 44), ovócitos cultivados em meio base mais 10% de soro inativado de cadela em estro. O percentual de ovócitos observados em metáfase I não indicou diferenças (P>0,05) entre T1 (2,1%) e T2 (0,0%), porém a taxa de ovócitos maduros (metáfase II) foi diferente (P<0,05), sendo 27,1% em T1 e 47,7% em T2. O mesmo fato ocorreu com a taxa de cromatina condensada (P<0,01), com 14,6 e 0,0%, respectivamente. Nos ovócitos sem configuração cromossômica, não foram observadas diferenças (P>0,05), sendo 56,3% em T1 e 52,3% em T2. Estes resultados indicam que a adição de soro de cadela em estro no meio de cultivo oferece melhores condições para o desenvolvimento in vitro, quando comparado à de soro de vaca em estro.

This study aimed to evaluate the effect of estrus on in vitro canine oocyte. A total of 92 oocyte from bitches under ovary-hysterectomy surgery was used. The oocytes were selected and randomly assigned to two different treatments, being T1 (n = 48) in vitro cultured for 96h using basic medium (TCM199 + 5µg/mL of LH + 20µg/mL of FSH), plus 10% of cow inactive serum in estrus and T2 (n = 44) basic medium plus 10% of bitch inactive serum in estrus. The percentage of oocyte observed on metaphase I do not indicate a difference (P>0.05) between T1 (2.1%) and T2 (0.0%). However, the rate of mature oocyte (metaphase II) was different (P<0.05), being 27.1% for T1 and 47.7% for T2. There was difference (P<0.05) in the condensed chromatin rate for T1 (14.6 %) and T2 (0.0 %), respectively. There was no difference (P>0.05) between T1 (56.3%) and T2 (52.3%) in oocyte with no chromosome configuration. These results indicate that supplementation with estrus bitch serum on culture media offer better conditions to in vitro development, when compared to estrus cow serum.

Avaliou-se a eficácia de duas soluções de manipulação (SM) de embriões de camundongas nos estádios de blastocisto inicial (Bin), mórula compacta grau I (McI) e II (McII), distribuídos aleatoriamente em três tratamentos (T), de acordo com a solução de manutenção. No T1 usou-se PBS modificado (controle); no T2, SME e no T3, SME enriquecida. Os embriões foram mantidos durante quatro horas na solução de manutenção e posteriormente classificados quanto ao estádio de desenvolvimento e à qualidade embrionária. Logo após, foram cultivados em meio TCM 199 e classificados novamente quanto ao estádio de desenvolvimento e à qualidade embrionária. A taxa de desenvolvimento dos embriões após manutenção por quatro horas em solução de manipulação foi menor (P<0,05) nos embriões do controle, comparada à de embriões do SME e SME enriquecida, diferença esta não observada (P>0,05) após o cultivo in vitro. Os embriões McII do T3 tiveram maior desenvolvimento (P<0,05) em relação aos embriões do T1 e T2, indicando o efeito benéfico do enriquecimento da solução SME. Conclui-se que as soluções de manipulação SME e SME enriquecida influenciaram beneficamente o desenvolvimento de embriões

The effect of embryo manipulation solution followed by in vitro culture in mice embryos was studied. The embryos at early blastocyst (Bin), and compact morula grades I (McI) and II (McII) were randomly assigned into three treatments. T1 used modified PBS (control), T2 used EMS, and T3 used EMS supplemented. In each treatment, the embryos were kept in manipulation solution for four hours. Finishing the manipulation period, the embryos were classified according the development stage and quality. Following, the embryos were cultured in TCM 199. After the culture period, the embryos were evaluated according to quality and development stage. The development rate for Bin, McI, and McII after maintenance for four hours in manipulation solution was lower for control embryo (P>0.05) as compared to EMS and EMS supplemented embryos. After in vitro culture, no differences (P>0.05) on embryo development rate among control, EMS, and EMS supplemented were observed. Moreover, McII from EMS supplemented had a higher development (P<0.05) (93%) as compared to control (82.5%) and EMS (83.9%), suggesting a beneficial effect of EMS supplemented. EMS and EMS supplemented embryos had a positive effect on embryo development, showing higher embryo development than those in PBS solution

A simplified, fast, and innovative method was developed to count the total cell number in blastocysts. Murine blastocysts (N = 195) were used in this study. They were obtained after 10h culture of initial blastocysts, compact morulae grades I and II recovered from superovulated mouse. After culture, the blastococysts were selected to test the new proposal of counting. The process was done after embryo fixation in a sodium citrate solution, and adherence in glass slide. Following, the coloration was done using a fast panoptic coloration kit. As a result, it was possible to identify the blastomeres and count them in each blastocyst. This method provided a fast and effective analysis of the total cell number when compared with other techniques. Moreover, this new method shows advantages related to the cell visualization, which can be done in more simple equipment like stereoscopic microscope. Other interesting observed point was the long period of time and quality that the coloration stays on slides, considering other techniques.