Effect of Trehalose and Natural Honeybee as Cryoprotectant on the Viability of Vitrified Thawed Matured Bovine Oocytes

Vitrification has been extensively used in cryobiology studies especially in oocytes studies. Each vitrification medium produces different vitrification capabilities depending on the vitrification medium components. Until today there is still a possibility of finding better vitrification medium by altering the sugar component in vitrification medium to find new superior combination. The objectives of this study are (1) To determine the effectiveness between Trehalose and Honeybee as cryoprotectants on viability of vitrified-thawed bovine oocytes (2) To determine the effectiveness of superstimulation protocol for ovarian follicular growth before Ovum Pick Up (OPU). Two cattle were subjected superstimulation protocol, per session conducted for total of five days where three days both cattle were administrated with 200 mg follicle stimulation hormone (FSH) within 24 h once and two days of “resting period” (FSH starvation) totalling for two session. The “coasting period” (FSH starvation) between first and second session were three days (72 h). OPU occurred at the fifth day (120 h) for oocytes collection. The ovarian growth was observed by ultrasonographic examination right before OPU. Prior
vitrifying oocytes by solid surface vitrification (SSV) with treatment Trehalose media (T1) and Honeybee media (T2) followed by warming protocol, oocytes subjected to in
vitro maturation (IVM). The oocytes viability were evaluated by fluorescein diacetate (FDA) staining. The follicular growth from first session were large size follicles for both
cattle but not the second session. Number of oocytes obtained from first session were (n=60) and second session (n=0). Oocytes viability for treatment Honeybee media (90.9%) was significantly higher Trehalose media (70.4%) at student t-test (p≥0.05). The presence study suggests the follicular growth are affected by FSH administration. The presence study also suggests the used of honeybee might be a superior alternative as sugar component in vitrification medium for vitrifying bovine oocytes.