Author/Authors :
Zeinoaldini، Saeed نويسنده Department of Animal Science, Faculty of Agricultural Science and Engineering, University of Tehran, Karaj, IR Iran , , Jafari، Zohre نويسنده Department of Animal Science, Faculty College of Agriculture and Natural Resources, University of Tehran, Karaj, IR Iran , , Sarmast، Faeze نويسنده School of Pharmacy, Shaheed Beheshti University of Medical Sciences, Tehran, IR Iran , , Torbati، Elham نويسنده Department of Microbiology, Islamic Azad University, Tehran Shomal Branch, Tehran, IR Iran , , Dadashpour Davachi، Navid نويسنده Department of Animal Sciences, College of Natural Resource and Agriculture Research, University of Tehran, Karaj, IR Iran ,
Abstract :
To current knowledge, different oocytes recovery methods have high impact on in vitro embryo production (IVEP). The aim of this study was to define an efficient recovery method for oocytes harvesting from slaughterhouse material, and its effect on IVEP yield. Ovaries were obtained from a local slaughterhouse and transported to laboratory in an insulate container physiological saline solution at 37 ºC. Oocytes were recovered via an aspiration pump (set at a flow rate of 10 mL H2O / min) with a disposable 20 G needle attached, oocyte recovery with centrifugation (ORC procedure), puncture and slicing. Oocytes were categorized into three classes (class I; oocytes with more than 3 layers of cumulus cell, class II less than 3 layers with damaged cumulus cells, and class III; denuded oocytes) and cultured in tissue culture medium (TCM) 199 medium for a period of 24 h in separate groups. Subsequently matured oocytes were subjected to in vitro fertilization (IVF) and afterwards, presumptive zygotes proceed to subsequent in vitro culture (IVC) for 6 days to complete blastocysts development. The present study showed that the oocyte recovery rate using ORC was significantly higher (P < 0.05) compared to other recovery methods. While no significant differences in the proportion of oocytes reaching M-II stage were recorded when using different oocyte recovery methods. Using aspiration for oocyte recovery led to lower proportion of class-I oocytes (60.0 ± 2.1) compared to ORC (82.0 ± 1.2), slicing (80.0 ± 2.1) and puncture (80.0 ± 1.5) methods. The results revealed that oocytes harvesting techniques are effective in the rate of cleavage and blastocysts development, and despite same meiotic resumption rate in all treatments, it would be better to use ORC for oocytes harvesting.
