Enhancement of in vitro maturation rate of oocytes in buffalo with a protein fraction obtained from the ovarian follicular fluid of buffalo

Ovarian follicular fluid is commonly being used as an input in the in vitro maturation (IVM) medium of oocytes. This study was conducted to find the oocyte stimulatory protein fractions of bubaline ovarian follicular fluid (buff). In this study the bubaline oocytes were studied. A protein fraction in IVM medium of bubaline oocytes was studied. A protein fraction precipitated from cell and steroid free buFF with ammonium sulphate at 0-25% saturation level was purified through sephadex-G 25 gel filtration column. There was no protein in the elution volume, but the void volume fraction consisted of 4 Proteins of Mr 30, 52, 65 and 205 kDa. Bubaline oocytes were matured in vitro with a basic culture medium consisting of TCM 199 + bovine serum albumin (0.3%) + gentamycin (50 mg/ml) at 38.5 degrees C, 95% relative humidity and 5% CO2 in air. When the protein fraction of buFF was incorporated in the basic lVM medium at different dose levels of 0, 5, 10 and 15 ng/ml of maturation medium, it stimulated the in vitro maturation rate of bubaline oocytes in a dose dependent manner. Hence, it was concluded that the stimulatory protein fraction of the bubaline ovarian follicular fluid can be supplemented in the IVM medium of bubaline oocytes at 20 ng/ml dose.