Effect of heparin on the capacitation of frozen – Thawed bull sperm used in the In-vitro fertilization of oocytes matured In-vitro

Abstract

Ovarian Oocytes were matured and fertilized in-vitro with frozen-thawed bull sperm capacitated in-vitro .Frozen – thawed sperm were swim up separated in a modified Ca⁺⁺ - free Tyrode’s medium and washed twice for 10 min. on each occasion. The washed sperm were then diluted ( 1:1 ) with same medium containing heparin so as to give final concentration of 0, 10, 50 and 100 ug/ml. Incubation of heparin treated sperm was carried out at 39C^˚ in 5% Co₂ for a periods of either 15 min. or 30 min. The inseminated Oocytes were cultured at 39C^ͦ  5% Co₂ in air for 20-48h and were then fixed, stained and examined for evidence of maturation and fertilization. The results show that a high percentage (96%) of Oocytes to reach metaphase II within 28 hours. It was also observed that the use of heparin was highly effective in enabling sperm to penetrate Oocytes. The proportion of Oocytes fertilized was increased significantly as the concentration of heparin in the capacitation medium increased significantly as the concentration of heparin in the capacitation medium increased ( 4.8%, 50.9%, 69.3%, and 83.6% for 0,10,50 and 100 µg/ml, respectively; P˂0.01 ). The percentage of Oocytes cultured with cow serum developed to 2-4-cells in a significantly greater than those cultured with BSA (63.3% VS 32.1%; P˂0.01).