| dc.description.abstract | This study was conducted with two objectives: Firstly, establish the IVM system by using dbcAMP to examine the meiotic competence of porcine oocytes. Secondly, to clarify interactions between oocytes and cumulus cells (CCs) on the level of oocytes cumulus expansion and oocyte maturation during IVM of cumulus-oocyte complexes (COCs) in a chemically medium using a system that allows to track and observe individual oocytes in in vitro maturation. The basic culture medium was bicarbonate-buffered TCM-199 supplemented with 10% (v/v) fetal bovine serum (GIBCO, life technologies, US), 0.1 mg/ml sodium pyruvate (sigma-Aldrich Co, LLC, japan), 10% (v/v) porcine follicular fluid, 0.1 IU/ml human chorionic gonadotropin (hCG) (Sigma, USA). Medium culture were divided to 2 types of different culture medium with adding 0.5mM( 0.5μL + 250μL of basic culture medium) and 1mM( 1μL + 250μL of basic culture medium) to reach metaphase II of oocytes after culture for maturation with 0.5mM of dbcAMP or 1mM of dbcAMP. Results of thesis indicates that dbcAMP is not capable of causing a meiotic arrest in collected oocytes because added of 1-mM dbcAMP in the medium during the first period on 16-18 hours of IVM but percentages of cumulus expansion was occupied more than 90%, althought comparing among three groups with no significantly differences of MII stages, but with these good oocytes expanded may be a signalling for establish IVF in next researching. I suggested supply 1mM concentration of dbcAMP in IVM will increase the ability of cumulus oocytes expansion. Beside, that is findings contribute to a better understanding of cAMP involvement to induce good quality of mature porcine oocyte and this maybe provide a basis to develop a culture system to obtain the higher normal embryo after fertilization.
Key word: Syndronized oocyte stages, cAMP oocytes, dibutylryl cyclic AMP, in vitro maturation. | en_US |
