USE OF DEAD CELL REMOVAL KIT FOR THE IMPROVEMENT OF RAM SEMEN QUALITY: SHORT COMMUNICATION
Keywords:
ram semen, CASA, MACS, dead cell removal, flow cytometryAbstract
The main objective of this preliminary study was to improve the ram semen quality by the removal of dead and apoptotic cells from the ejaculates. For this purpose, ram spermatozoa were incubated with the Dead Cell Removal kit and magnetically sorted using two procedures with different sample loading rate (Deplete and Depletes) by a fully automated cell sorter. Fresh semen samples (control) as well as both sorted fractions (negative and positive) were analysed for motility parameters using CASA and for the proportion of dead cells using flow cytometry. As expected, a significant increase (P < 0.05) in the number of dead cells and decrease (P < 0.05) in the spermatozoa motility were observed in the positive fractions when sorted by both procedures. However, the viability of negatively sorted spermatozoa was not improved and their motility was insignificantly decreased. In conclusion, although the presented study demonstrates the possible use of a MACS technique for the elimination of ram spermatozoa with compromised membrane, the chosen sorting procedures seem to be insufficient to obtain high purity of spermatozoa with intact membranes. More sensitive depletion programmes should be tested in further studies.
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