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Free Content Elimination of Apoptotic Spermatozoa from Rabbit Insemination Dose Using Annexin V Associated with the MACS Technique. A Preliminary Study

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Abstract:

The aim of this study was to verify whether the separation and elimination of the apoptotic fraction in rabbit semen using a MACS technique may improve sperm fertility potential and consequently rabbit kindling rate. Semen samples from 25 New Zealand White (NZW) rabbit males were collected using an artificial vagina and evaluated using the CASA system for concentration and motility. For artificial insemination the best 11 bucks were chosen based on motility parameters. Their ejaculates were mixed to make a heterospermic pool and routinely diluted in a commercial insemination diluent (MiniTüb, Tiefenbach, Germany) at a ratio of 1:6. Diluted heterospermic spermatozoa were filtered through a Sartorius filter to wash out seminal plasma, re-diluted in binding buffer (Annexin V Microbead Kit, Miltenyi Biotec, Germany) at a ratio of 1:3.66 and divided into two groups: an experimental group intended for MACS separation and control group without MACS separation. Then hormonally treated females of NZW rabbits were inseminated with fresh doses of filtered heterospermic semen (n=27; 0.5 ml I.D. per female) and MACS separated semen (n=28; 0.5 ml I.D. per female). Separation and subsequent elimination of apoptotic spermatozoa (positive selection) from the insemination dose (after negative MACS selection) was verified under in vivo conditions on the basis of increased kindling rate in the experimental group in comparison with kindling rate in the control group (81.3% vs. 73.8%). In conclusion, elimination of apoptotic spermatozoa by the use of the MACS technique results in a slight improvement in kindling rate of rabbit does.

Keywords: ANNEXIN V; APOPTOSIS; KINDLING RATE; MAGNETIC-ACTIVATED CELL SORTING; RABBIT; SPERMATOZOA

Document Type: Short Communication

DOI: https://doi.org/10.3409/fb59_1-2.65-69

Publication date: 2011-01-01

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  • Folia biologica is an international quarterly journal that publishes papers on the broad field of experimental zoology, nuclear and chromosome research, and also ultrastructural studies. All papers are subject to peer reviews. Indexed in: ISI Master Journal List, Current Contents, Polish Scientific Journals Contents. I.F. 0.667
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