The role of tubal fluid culture conditiion during oocyte maturation and arrest

Maturation of oocyte started at embryonic stage, it hesitated during prohase I of meiosis, started to develop in puberty with developing follicules. During the development stages of oocyte at meiosis I and meiosis II, many molecular mechanisms controls. These mechanisms also provided to transition from one phase to another phase of oocyte and was complemented by the development of mature oocytes for fertilization. Oocyte maturation problems and/or pauses of development of oocyte during meiotic stage (remain in arrest) could not be seen fertilization. Therefore, the culture condition for oocyte had to be develop. Provided appropriate culture media and to protect the viability of embryos cultured in vivo culture system that could mimic conditions were needed as much as possible. For these purpose, in addition to IVF culture medium, co-culture systems might be preferred. According to these data, in our study, we investigated to analyze the effect of tubal fluid culture system on oocyte maturation and embryo development potentials. For this purpose, after superovulation of mouse, meiosis I and meiosis II stage of oocytes would be cultured in culture condition. In the first group, oocytes with cumulus cells would be at stage of meiosis I, in the second group oocytes without cumulus cells would be at stage of meiosis II would be culture in medium with tubal fluid. A total of 6 groups would be examined. Groups: Group 1 : Metaphase I oocyte culture Group 2 : Metaphase I oocyte culture + tubal fluid Group 3 : Metaphase II oocyte culture Group 4 : Metaphase II oocyte culture + tubal fluid After 48 hour of culture, oocytes would be evaluated anti-cyclin B, anti-MPF and anti-MOS distribution using whole mount immunofleurosans staining technique. Apoptotic cells by TUNEL assay would be evamined with the same time. In addition, to analyze secreted anti-cyclin B, anti-MPF and anti-MOS distribution during ovarian cycle, ovarian tissue would also be collected (form 5 mouse) and after routine paraffin embedding protocols, the sections would be stained to with anti-cyclin B, anti-MPF and anti-MOS using immunoperoxidase technique. In ovarian tissue sections, the apoptotic cells would be analyze using TUNEL assay. While the oocytes taken during meisis I were observed to pass to phase of meiosis II following the culturing in the tubal fluid ,the oocytes in meiosis II were noted to preserve their vitality in the tubal fluid culture and especially during both phases with the secretion of Emi.