Methods for equine preantral follicles isolation: quantitative aspects.

The study conducted research on various methods of isolating preantral follicles in horse ovaries. The main finding showed that mechanical methods proved more effective than combined mechanical-enzymatic processes, and both saline solution (SS) and phosphate-buffered saline (PBS) were viable for equine preantral follicle preparation.

Objective and methodology

• The main objective of the study was to investigate and compare the effectiveness of different methods for the isolation of preantral follicles (PAFs) in horse ovaries. These included mechanical and combined mechanical-enzymatic procedures, as well as the use of saline solution (SS) and phosphate-buffered saline (PBS).
• To do this, horse ovaries were subjected to both the mechanical isolation method using a mixer and combined mechanical and enzymatic digestion using a collagenase. This was done for incubation times of both 10 and 20 minutes.

Results

• In the first group, an average of 4.1 to 4.9 PAFs were collected using the mechanical-enzymatic method, compared to 71.1 to 19.2 when using the purely mechanical procedure. This showed a significant difference between the two methods favoring the mechanical procedure.
• When using SS and PBS as media for equine PAFs, the yields were 35.7 to 34.3 and 39.6 to 39.6 respectively, showing no significant difference between the two solutions.
• In the second group, using an incubation time of 20 minutes, yielded similar results, with the mechanical method again harvesting more PAFs (63.2 to 22.9), compared to the mechanical-enzymatic method (7.1 to 10.6).
• Using SS and PBS in the second group again resulted in similar amounts of PAFs being harvested (35.5 to 36.4 and 34.9 to 31.1 respectively).

Conclusion

• The findings showed that the mechanical method of isolating equine PAFs was significantly more effective than the combined mechanical-enzymatic method.
• Additionally, the use of both saline solution (SS) and phosphate-buffered saline (PBS) were validated as effective media for the preparation and potential manipulation of equine PAFs.