الفهرس 
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Abstract
There are numerous factors that may influence the efficiency of IVF protocol during different protocol steps (in-vitro maturation, in-vitro fertilization or in-vitro culture).
The present study was designed to investigate the efficiency of IVF protocol after fertilization of COCs or denuded oocytes under different conditions as different sperm-oocyte co-incubation time (6, 12 and 18 hrs), using sperm cells from different bulls (3 bulls) or addition of specific additives to fertilization medium (heparin, theophylline and penicillamine hypotaurine epinephrine mixture). Additionally, we investigated the interaction between all these conditions with the type of the oocytes (COCs vs denuded)
Buffalo ovaries were collected within 0.5 h after slaughtering and transferred to Lab.in sterile saline with the antibiotic in a thermos container. Cumulus oocytes complexes (COCs) were aspirated from all antral follicles. Oocytes with homogenous cytoplasm and surrounded with 2 or more layers of compact cumulus were matured in-vitro for 24 hrs at 38.5 ºC and under 5% CO2 in humidified.
In all experiments, some of the in-vitro matured oocytes were used as COCs and the others were denuded through vortex in supplementation with 10 Iu/ml hyaluronidase.
In first experiment COCs and denuded oocytes were incubated with the sperm cells for 6, 12 or 18 hrs. In second experiment COCs and denuded oocytes were fertilized with.