In vitro maturation, fertilization and development of buffalo oocytes

In Egypt, buffaloes are considered the main dairy animals and have a vital economic interest, however, this species is mostly out of the attention and still until now obscure. Currenteconormc demand for buffalo milk and meat has dictated breeding strategies which necessitate the extension of modern biotech- neology of embryo transfer and in vitro fertilization to this species. The in vitro fertilization of buffalo oocytes represents oneed further substantial improvement. The present work aimed to develop a reliable in vitro technique for buffalo oocyte maturation, fertilization and culture, that could regularly provide a large number of normally fertilized oocytes for further biotechnapplication.The oocytes were harvested from the buffalo ovaries post slaughter of the buffaloes from all visible surface immature follicles ranged from 2-8mm in diameter. The oocytes were washed threetimes with TCM-199 medium supplemented with sodium pyruvate and gentamycin sulfate. Only oocytes that surrounded by compact multi layers of cumulus cells, with evenly granular ooplasm wasselected for in vitro maturation. The good quality oocytes were matured in TCM-199 that supplemented with 50J.lg/ml sodium pyruvate, 1 % (v/v) antibiotic antimycotic solution and 10% FCS, 5J.lg/ml, FSH, 10j.lg/ml LH and 1 j.lg/ml estradiol -17B. The rnediwas sterilized by filtration through 0.22 Millipore filter membrane.The selected oocytes were covered by sterile paraffin oil and