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Fluorimetric Techniques for the Assessment of Sperm Membranes
08:58

Fluorimetric Techniques for the Assessment of Sperm Membranes

Published on: November 28, 2018

Deterioration of plasma membrane is associated with activated caspases in human spermatozoa

Uwe Paasch1, Sonja Grunewald, Guenter Fitzl

  • 1Department of Dermatology/Andrology Unit, University of Leipzig, Leipzig, Germany.

Journal of Andrology
|March 14, 2003
PubMed

Insights

Magnetic-activated cell sorting (MACS) effectively separates sperm with damaged membranes, identified by annexin V binding. This method significantly reduces activated caspases (aCPs), improving sperm quality in both donors and infertile patients.

Area of Science:

  • Reproductive biology
  • Cell biology
  • Biotechnology

Background:

  • Sperm plasma membrane integrity is crucial for male fertility.
  • Deteriorated sperm membranes exhibit phosphatidylserine (PS) externalization.
  • Activated caspases (aCPs) are indicators of sperm cell damage and apoptosis.

Purpose of the Study:

  • To evaluate the efficacy of magnetic-activated cell sorting (MACS) using annexin V-conjugated microbeads (ANMBs) for separating spermatozoa with compromised plasma membranes.
  • To assess the levels of activated caspases (aCPs) in separated sperm fractions.
  • To compare sperm quality between fertile donors and infertile patients.

Main Methods:

  • Semen samples were fractionated using annexin V-MACS.
  • Levels of bound annexin V and activated caspases (aCPs) were quantified.
  • Flow cytometry and electron microscopy were used to confirm separation efficiency.

Main Results:

  • Spermatozoa from infertile patients showed significantly higher levels of bound annexin V and aCPs compared to donors.
  • Annexin V-MACS significantly reduced the percentage of spermatozoa with aCPs in both donor and infertile groups.
  • The ANMB-negative fraction exhibited a higher quality spermatozoal population.

Conclusions:

  • ANMB-MACS is an effective technique for removing spermatozoa with externalized PS and reduced quality.
  • Elevated aCPs are indicative of deteriorated sperm membranes.
  • This method offers a promising approach to improve sperm quality for assisted reproductive technologies.

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