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Published on: January 14, 2021
Evaluation of sperm recovery following annexin V magnetic-activated cell sorting separation
Tamer M Said1, Ashok Agarwal, Sonja Grunewald
1Centre for Advanced Research in Human Reproduction, Infertility and Sexual Function, Glickman Urological Institute and Department of Obstetrics-Gynecology, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH 44195, USA. tamersaid@repromedltd.com
Abstract:
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. Following MACS, the percentage of cells collected in the annexin-negative fraction was significantly higher than the annexin-positive fraction and the sperm recovery rate was 73.8 +/- 12.1%. In conclusion, the integration of MACS with DGC can be considered as an effective sperm preparation technique that does not lead to significant cell loss. Separating a distinctive population of non-apoptotic spermatozoa with intact membranes may optimize the outcome of assisted reproduction.
Insights
Magnetic-activated cell sorting (MACS) effectively prepares sperm by removing apoptotic cells. This method, combined with density gradient centrifugation (DGC), ensures high sperm recovery rates and preserves membrane integrity for assisted reproduction.
Area of Science:
- Reproductive Biology
- Cell Biology
- Biotechnology
Background:
- Externalized phosphatidylserine is a marker of apoptosis in spermatozoa.
- Magnetic-activated cell sorting (MACS) can identify and remove apoptotic cells.
Purpose of the Study:
- To evaluate sperm recovery rates when using MACS as a sperm preparation technique.
- To assess the efficacy of MACS in conjunction with density gradient centrifugation (DGC).
Main Methods:
- Spermatozoa were prepared using either DGC combined with MACS or DGC alone.
- Paramagnetic annexin V-conjugated microbeads were used for MACS to target cells with externalized phosphatidylserine.
- Sperm recovery was quantified and compared between the two preparation groups.
Main Results:
- The annexin-negative fraction (non-apoptotic cells) contained a significantly higher percentage of cells compared to the annexin-positive fraction.
- The overall sperm recovery rate following MACS was 73.8 +/- 12.1%.
- No significant cell loss was observed when MACS was integrated with DGC.
Conclusions:
- Integrating MACS with DGC is an effective sperm preparation method.
- This technique allows for the separation of non-apoptotic spermatozoa with intact membranes.
- Optimizing sperm selection through MACS may improve assisted reproduction outcomes.
