Pronuclei formation subsequent to intracytoplasmic sperm injection in bovine

Abstract

ICSI represents the top of the range of the assembly protocols of assisted reproduction which presumes the existence of sophisticated equipments, prior acquired routine in preparing the oocytes and sperm and a very good organization and synchronization of the working time. The aim of the present paper is represented by the assembly ICSI technique in the assisted reproduction laboratory within CLC-HC Timisoara . 70% of the oocytes were considered able for ICSI and out of this number 12 (17%) were destroyed meanwhile micromanipulation; the remaining were fertilized using sperm treated with PVP (G1), TritonX (G2) or untreated sperm (control). Taking into account the outcome results the superiority of the method that prepares the sperm with Triton X, even though the fertilization percentage (35%) is clearly inferior toward the ones reported (60-80% by Sekhavati et al., 2012). Using PVP to prepare the sperm has generated a lower percentage of success (30%) besides a overwhelming proportion of oocytes with one pronucleus (1PN-80%) or unfertilized (NE-20%). Previous reports emphasize the fact that the volume of PVP which gets in the oocyte cytoplasm consecutively injecting the sperm can have a harmful effect on the zygote. The efficiency of Triton X to remove the acrosome, in this way dislodging a consistent enzymatic volume and allowing to decondense the male pronucleus it is demonstrated by the 7 fertilized oocytes (35%) by injecting the sperm treated previous with this solution. Untreated sperm it has generated the oocytes fertilization only in proportion of 11% whereas the male pronucleus does not cover in an useful time and requested proportion the transformations needed to support the fecundation. The culture media used for gamete and zygote preparation had generated lower results toward the reports in the literature. The modest outcomes can be explained both through the culture media composition and through the long execution of ICSI. The presence of both pronuclei in some of the oocytes subdued to ICSI (between 30 and 35% depending on the substance used to treat the sperm), proves our capacity to assemble ICSI technique in the assisted reproduction laboratory within CLC-HC.