Isolation and characterization of preantral follicles from fetal bovine ovaries

A simple, mechanical method is described for the isolation of preantral follicles bovine foetuses of 220-280 days of gestation. On average, 2918+621 (s.d.) preantral follicles were isolated per ovary. The isolated preantral follicles were characterized on the basis of the morphological appearance of the surrounding granulosa cells, the number of granulosa cell layers, and their diameter. The results show that primordial, primary, and secondary follicles differ morphologically and that they can be classified by their diameter

DESENVOLVIMENTO DE FOLÍCULOS PRÉ-ANTRAIS BOVINOS IN VITRO EM MONOCAMADA DE CÉLULAS OVARIANAS IN VITRO DEVELOPMENT OF BOVINE PREANTRAL FOLLICLES IN MONOLAYER OF OVARIAN CELLS

O presente trabalho teve como objetivos determinar a influência de células ovarianas no desenvolvimento in vitro de folículos pré-antrais, avaliar a viabilidade das células ovarianas em monocamada e a influência do soro na manutenção de folículos pré-antrais in vitro. Folículos pré-antrais (FPs) e células ovarianas foram isolados de ovários de fetos bovinos, com idade entre 6 e 8 meses de gestação, oriundos de matadouro. Células ovarianas em monocamada foram cultivadas em meio TCM-199, e a viabilidade celular, após o cultivo na presença ou ausência de FSH, foi determinada com o corante vital azul de tripan. FPs foram distribuidos em quatro tratamentos e cultivados em TCM-199 modificado, contendo soro de novilho castrado (SNC), SNC em monocamada de células ovarianas (MCO), MCO com FSH ou meio definido com álcool polivinílico (PVA) como macromolécula. A viabilidade celular não foi afetada em conseqüência da presença ou ausência de FSH. No entanto, houve um incremento significativo no tamanho dos FPs cultivados na presença de SNC, MCO e FSH (P<0,05), o que não foi observado nos demais tratamentos. A adição de FSH ao meio de cultivo não influencia a viabilidade de células ovarianas ao longo de 20 dias. Folículos pré-antrais, com área superficial entre 4000 e 8000mim², mas não folículos com área superficial superiores a 8000mim², crescem in vitro na presença de células ovarianas, independentemente da presença de FSH.<br>The aim of the present work was to determine the influence of ovarian cells in the in vitro development of preantral follicles (PF). The viability of monolayer ovarian cells and the effect of the serum in the survive of in vitro PF was also investigated. Ovarian cells and PF were isolated from ovaries of bovine fetus between 6 and 8 months of pregnancy, obtained in a slaughterhouse. Monolayer of ovarian cells were cultured in a modified TCM-199 in the presence and absence of FSH and its viability after incubation was determined with Trypan Blue. PFs were divided in four different treatments, cultured in modified TCM-199, containing serum of castrated steer (SCS), SCS in monolayer of ovarian cells (MOC), MOC with FSH or a defined medium with polyvinyl alcohol (PVA) as macromolecule. The cellular viability was not affected by the presence or absence of FSH. However, PFs had a significant development (P<0.05) when cultivated in the presence of SNC, MCO and FSH, which was not observed in the other treatments. With these results, it was possible to conclude that FSH does not influence the viability of the ovarian cells when cultured in a monolayer for 20 days. Preantral follicles, measuring between 4000 and 8000mum² but not with size greater than 8000mum², grow on ovarian cells in vitro, independently of FSH

Mechanical isolation of capuchin monkey (Cebus apella) preantral ovarian follicles Isolamento mecânico de folículos ovarianos pré-antrais de macaca-prego (Cebus apella)

The aim of this study was to adapt a mechanical procedure for the isolation of intact preantral follicles from Cebus apella ovaries. The interval effect of serial sections of the tissue chopper was tested on a number of preantral follicles isolated from ovaries (n=6) of three C. apella females, two prepubertal and one adult. Ovaries were divided into four equal parts and fragmented with a tissue chopper, adjusted for serial sections at intervals of 250, 500, 750 and 1,000µm, respectively. Isolated follicles were counted in a Neubauer's chamber and classified as primordial, primary or secondary. The number (mean±SE) of preantral follicles isolated from 1/4 ovary varied from 68,330+17,590 (at the 1,000µm cut interval) to 300,830+111,460 (at the 500µm cut interval. The mean diameter of the isolated preantral follicles varied from 11.6µm to 27.8µm.<br>O presente trabalho objetivou adaptar um procedimento mecânico para o isolamento de folículos pré-antrais a partir de ovários de Cebus apella. Para isso, foi testado o efeito do intervalo de cortes seriados do tissue chopper sobre o número de folículos pré-antrais isolados a partir de ovários (n=6) de três fêmeas de C. apella, duas pré-púberes e uma adulta. Os ovários foram divididos em quatro partes iguais e fragmentados com auxílio de um tissue chopper, ajustado para a realização de secções seriadas a intervalos de 250, 500, 750 e 1000µm, respectivamente. Os folículos isolados foram contados em câmara de Neubauer e classificados em primordiais, primários e secundários. O número (média ± EP) de folículos pré-antrais isolados de 1/4 de ovário variou de 68.330+17.590, no intervalo de corte de 1.000µm, a 300.830+111.460, no intervalo de corte de 500µm, o de melhores resultados. O diâmetro médio dos folículos pré-antrais isolados variou de 11,6µm a 27, 8µm

The embryonic development of the bovine stomach revisited.

The adult anatomy and physiology of the bovine (Bos taurus) stomach have been investigated extensively. Despite the many studies, however, the early development of the stomach has not yet been fully elucidated. The goal of the present study, therefore, was to review the available literature, to visualize the embryonic and early foetal development of the bovine stomach and to shed light on unresolved issues. The stomachs of fifteen bovine embryos and eleven foetuses from 26 to 80 days of gestation were photographed both in situ and after exenteration and critical point drying. A series of photographs was obtained that yielded a contiguous and comprehensive view of all the developmental changes that occurred until the virtually final configuration of the stomach was attained. In addition, the serosal surface was studied by electron microscopy, thus revealing subtle regional differences in the lining of the peritoneal cavity. Our observations corroborate the contention that all the compartments evolve from the fusiform primordium and that no outgrowth at the level of the oesophagus occurs. The greater curvature as well as the attachment line of the dorsal mesogastrium shift to the left, which is similar to the process in monogastrians. The rumen and reticulum develop from separate protrusions, and further compartmentalization results from constrictions and bulges and not from folding. Between 55 and 60 days of gestation, the entire bovine stomach except for the abomasum eventually relocates to its final position. In summary, previously debated key issues were addressed and integrated with current findings