الفهرس 
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Abstract
In Egypt, buffaloes are considered the main dairy animals and have vital economic interest; however, this species is mostly out of the attention and still until now obscure. Current economic demond for buffalo milk and meat has dicatd breeding strategies which necessitate the extension of modern biotechnology of embryo transfer and in vitro fertilization to this species.
The in vitro oocyte maturation, fertilization and culture are sub optimal and need further substantial improvement. This study was carried out to improve maturation rate of buffalo oocytes by investigating the effect of recovery methods of buffalo oocytes on recovery rate and quality of buffalo occytes. The effect of different culture media on IVF , fertilization and culture rates of buffalo occytes as well as supplementation of IVM media with growth factor as (IGF-1) , antioxidant as (GSH and cystine) and amino acids as (L-Tyrosine) to improveIVM, fertilization and in vitro cleavage rates of buffalo oocytes.
A total of 300 ovaries were collected. Oocytes were aspirated or sliced and the number of recovered oocytes was counted per trial and the number of oocytes / ovary was calculated. The recovery rate was determined. Oocytes were classified according to the appearance of cumulus cells into excellent, good, fair and poor.
They were cultured either in TCM-199, SOF, KSOM or Ferticult with different additives (IGF-1, GSH and Cystine or Tyrosine) with different concentrations for 24 h in CO2 incubator at 38 oC.
Matured oocytes were fertilized in vitro by frozen thawed semen, capacitated in vitro by heparin and sodium caffeine benzoate, at a final concentration of 2x 106 sperm/ml. The sperm oocytes were incubated for 22 hr in TALP medium. Then cultured in C-SOF medium with 10% FCS and incubated in CO2 incubator at 38 oC.
The present study revealed that the Quality and number of oocytes recovered from buffalo ovarian follicles affected by recovery oocyte technique and aspiration technique best than slicing of buffalo ovary. Also, In vitro maturation , in vitro fertilization and in vitro culture of buffalo oocytes was affected by maturation medium. SOF and Ferticult medium enhanced cumulus cell expansion , nuclear maturation , fertilization and cleavage rates more than TCM-199 or KSOM media and The IVM, IVF and IVC rates of buffalo oocytes was affected by additives on maturation medium as GHS, IGF-1, L-Tyrosine.