Selecting motile, non-apoptotic and induced spermatozoa for capacitation without centrifuging by MACS-Up method.

Abstract

In newly improved MACS-Up method, magnetic field has been applied to separate non-apoptotic spermatozoa directly from the neat semen. The spermatozoa during passing through a viscous layer, located on the neat semen, contacted with progesterone and induced for the capacitation. Then, a clean population of non-apoptotic, and capacitated spermatozoa were selected in the pure culture media. Selected spermatozoa may be useful for use in ART. The 80semen samples from normozoospermic individuals were divided separately into 4 attempts. Semen analysis, SCSA (sperm chromatin structure assay), FLICA (fluorescein-labelled inhibitors of caspase) methods, immunoassay of phosphorylation of tyrosine residues of sperm proteins, nuclear DNA integrity, caspase 3 activity and sperm capacitation rate were all performed for evaluation of sperm parameters respectively. To examine all aspects, the MACS-Up method compared with DGC (density gradient centrifuging) and MACS-DGC methods separately. This method can isolate non-apoptotic spermatozoa directly from the neat semen, which has similar performance compared to the MACS-DGC method. Movement and passing spermatozoa through the viscous layer, and contact with progesterone, significantly induced spermatozoa for capacitation compared with the control group. Also, the MACS-Up in comparison with routine DGC method could select spermatozoa with significantly higher total and progressive motility, DNA integrity, induced sperm population for capacitation and normal morphology. MACS-Up can be developed as an effective, short-time, and ease of performing method and used practically to select functional spermatozoa as novel sperm selection procedure. However, for clinical use of MACS-Up, all clinical aspects of this method should be considered and evaluated.