Thesis (M.Sc.) - Cairo University - Faculty of Agriculture - Department of Animal Production

The current study was conducted to investigate the possible protective effects of curcumin supplementation on buffalo granulosa cells (GCs) under in vitro culture conditions. Buffalo ovaries were collected from local abattoir in physiological saline solution at 37oc and transported directly to laboratory. Follicular fluid containing GCs and cumulus-oocyte-complexes were aspirated from antral follicles with diameter 2-8 mm. The collected GCs were seeded (Approximately 375,000 viable cells) in a 8-well culture plate containing 199-tissue culture medium (TCM) and kept at 37{u00B0}C in a humidified atmosphere of 5% CO2. The curcumin was supplemented to TCM media at levels of 1, 2.5, 5 and 10 oM or kept without treatment as control group for 48 hours. The viability of cells was determined using the trypan blue test. Intracellular reactive oxygen species (ROS) level was assessed by measuring the fluorescent intensity of 6-carboxy-2{u2032}, 7{u2032}-dichlorodihydro fluorescein diacetate (H2DCFDA). In addition, mitochondrial activity of GCs was using Mitrocker Red Fluorescent stain. The results of the present study indicated that the viabilityof GCsunder culture conditions were significantly decreased p {u2264} 0.05 in groups treated with 1, 2.5, 5 and 10 oM curcumin (86.0%, 86.26%, 83.0% and 74.0%, respectively) compared to control group (93.60%).The two groups of granulosa cells cultured with 2.5 and 5 oM curcumin recorded greater levels of mitochondrial activity than the groups cultured with 1 oM and 10 oM curcumin. In addition, there was a significant increase in ROS level in group cultured with 10oM curcumin, compared to control and other experimental groups

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