Effect of polyvinylpyrrolidone (PVP) and growth factor combinations on in vitro growth factor of pig oocyte

Abstract

The purpose of this project was to evaluate the effects of polyvinylpyrrolidone (PVP) and growth factor, especially is androstenedione combinations for growing oocytes from antral follicles (2-3mm) of porcine oocytes to become fully grown oocytes. Moreover, meiotic competence had ability reach to matured oocytes and metaphase II by in vitro maturation after in vitro growth. In this study, growing porcine oocytes were collected from preantral follicles (2-3mm in diameter). Then those oocytes were cultured for 24 hours and 20 hours to determine the optimal duration, in vitro growth (IVG) medium which contained Dulbecco’s modified Eagle’s medium (DMEM), 0.01 IU/mL follicle-stimulating hormone (FSH), 1 mM cyclic adenosine monophosphate (cAMP), 0.5 μg/mL Estradiol-17β, 5% Fetal bovine serum (FBS), 5% porcine follicular fluid (pFF), 0.1mg/mL Sodium pyruvate (basic medium) and added 0.1 μg/ml androstenedione alone or combinated with 2% polyvinylpyrrolidone (PVP) by floating drop method and 96-well method. After in vitro growth cultured in three kinds of medium, growing oocytes could obtain fully grown stage at germinal vesicle I (GVI) (47.9%, 53.6%, 44.4%) while non-culture growing oocytes only had 12.9%. Then growing oocytes after IVG and IVM obtained meiotic competence and reach to germinal vesicle breakdown similarly with fully growth oocytes in three kinds of medium. Besides, IVG oocytes in medium 1 and medium 3 underwent IVM reached metaphase II similarly with fully grown oocytes (47.4% and 48.1% in medium 1 and 3; 42.6% of fully grown oocytes). Growing oocyte with combination of androstenedione and polyvinylpyrrolidone during IVG could obtain meiotic competence reach to metaphase II similarly with fully growth oocytes after IVM. Key words: androstenedione, polyvinylpyrrolidone, preantral follicles, porcine growing oocytes, in vitro growth, in vitro maturation