Stability of fluorochrome based assays to measure subcellular sperm functions.

Aim: To evaluate the long-term stability of the fluorescence signals of new fluorescence-based semen analysis assays for clinical application.

Methods: Semen samples from 87 unselected infertile patients were used to perform the following assays: (i) detection of active caspase-3 (n=17); (ii) integrity of the mitochondrial membrane potential (MMP) (n=17); (iii) externalization of phosphatidylserine (EPS) (n=16); and (iv) detection of intact acrosomes via CD46 (n=37). After the assays, 4% paraformaldehyde was added to all aliquots.

Molecular glass wool filtration as a new tool for sperm preparation.

Background: Magnetic-activated cell sorting (MACS) using annexin V-conjugated microbeads in a liquid phase eliminates apoptotic spermatozoa based on the externalization of phosphatidylserine (EPS) residues. The procedure allows the enrichment of a sperm population free of apoptosis markers, giving higher fertilization potential. Our aim was to determine if the annexin V binding principle can be transferred onto a glass wool filter system in order to produce a solid phase filter.

Evaluation of sperm recovery following annexin V magnetic-activated cell sorting separation.

Magnetic-activated cell sorting (MACS) using paramagnetic annexin V-conjugated microbeads eliminates spermatozoa with externalized phosphatidylserine, which is considered one of the features of apoptosis. The objective of this study was to evaluate sperm recovery following the use of MACS as a sperm preparation technique. Mature spermatozoa were separated and divided into two fractions: the first was prepared by density gradient centrifugation (DGC) and MACS, while the second was prepared by DGC only.

Magnetic-activated cell sorting before cryopreservation preserves mitochondrial integrity in human spermatozoa.

Superparamagnetic annexin-V conjugated microbeads are able to eliminate spermatozoa with externalized phosphatidylserine, a membrane feature of apoptotic cells as well as spermatozoa with deteriorated plasma membrane. Our objective was to evaluate the effects of annexin-V Magnetic-Activated Cell Sorting (MACS) in cryopreservation-thawing protocols and on integrity of sperm mitochondrial transmembrane potential and mitochondrial integrity survival rate (MSR). Mature spermatozoa of 10 healthy donors were prepared by density gradient centrifugation and divided into 2 aliquots afterwards.

Selection of nonapoptotic spermatozoa as a new tool for enhancing assisted reproduction outcomes: an in vitro model.

Magnetic cell sorting (MACS) using annexin V-conjugated microbeads eliminates apoptotic spermatozoa based on the externalization of phosphatidylserine residues. The procedure delivers two sperm fractions: annexin V-negative (nonapoptotic) and annexin V-positive (apoptotic). Our aim was to determine whether the sperm fertilizing potential can be improved by selecting a nonapoptotic fraction using MACS.

Effects of magnetic-activated cell sorting on sperm motility and cryosurvival rates.

Objective: To evaluate the effect of magnetic-activated cell sorting in cryopreservation-thawing protocols on sperm motility and cryosurvival rate.

Design: Prospective-controlled study.

Setting: Andrology department at a university-based medical institution.