Different Harvesting Techniques Used in Ovine in vitro Embryo Production
Scimetr: March 31, 2013, 1 (1); e87326
August 27, 2013
Article Type: Research Article
April 18, 2013
July 05, 2013
Z , Sarmast
E, Dadashpour Davachi
N. Different Harvesting Techniques Used in Ovine in vitro Embryo Production,
Background: To current knowledge, different oocytes recovery methods have high impact on in vitro embryo production (IVEP). Objectives: The aim of this study was to define an efficient recovery method for oocytes harvesting from slaughterhouse material, and its
effect on IVEP yield.
Materials and Methods: 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.
Results: 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.
Conclusions: 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.