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Title: Development of porcine preantral follicles in vitro
Author: Bark, Monica Clare
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2004
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The development of a culture system to sustain preantral follicle growth to a stage where fertilisable oocytes can be obtained remains elusive in the domestic species. The objectives of this thesis were to develop a culture system capable of supporting porcine preantral follicles, and identify possible markers of oocyte and follicle development. Follicles were cultured in two systems; individually in McCoys medium to identify the effects of ascorbic acid, FSH, and serum, and in NCSU medium individually (96-well plates) or in-groups (24-well plates) to identify the effects of follicle interactions. Results from the McCoys experiment revealed that somatic cell death was significantly reduced in follicles cultured in the presence of ascorbic acid in comparison with other treatment groups, but the vitamin was found to have no effect on follicle growth. Follicle growth was significantly enhanced by the addition of serum and FSH to serum-free medium, but FSH had no effect as a survival factor on granulosa cell death in follicles. Culture in NCSU medium revealed that follicles grew best when cultured individually in the presence of serum in 96-well plates in comparison to follicles cultured in-groups in 24-well plates. All other parameters of follicle health and development were found to be no different between follicles cultured in serum in-groups of individually. The identification of markers of development for follicles and oocytes could also aid the development of a culture system for preantral follicles. GDF-9 is a possible indicator of follicle and oocyte developmental stage. It has been identified in several species, including human, rodents, and domestic species, but not in the pig. In this study it was isolated using human and mouse primers, and sequenced. It was found to display 88% homology to the human sequence. RT-PCR revealed that it appears to be expressed strongly in the porcine oocyte, but not in granulosa, skin or intestinal cells. BMP-15 was also found to be oocyte-specific in the pig, using sheep primers to identify its location.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available