Isolation and cryopreservation of functionally competent equine leucocytes.

The research paper aims to establish a method for cryopreserving functionally competent neutrophils, a type of white cell, for use in endometritis therapy in horses. The process extracts leucocytes from the blood of healthy mares, freezes them in a special solution and demonstrates they remain viable and efficient after thawing.

Objective and Importance

• The primary intent of this study was to develop an effective method for the preservation of functionally competent neutrophils, which are crucial in treating equine endometritis, a common reproductive disorder in mares.
• The success of this method could significantly improve the effectiveness of endometritis therapy.

Methods Used

• The research used hypotonic lysis to separate leucocytes from whole blood extracted from healthy mares, a procedure which proved more effective than dextrose sedimentation.
• The separated leucocytes were then suspended in a cryoprotective solution composed of equine plasma with 5% dimethyl sulphoxide (DMSO).
• The frozen leucocytes were stored in liquid nitrogen, with a specific temperature gradient and a low cooling velocity.

Results

• The method resulted in the highest numbers of viable cells after defrosting, compared to other approaches.
• The defrosted PMNs retained high phagocytic capacity for Streptococci bacteria and were able to generate reactive oxygen species (ROS) after stimulation with a phorbol ester.
• The competency of these defrosted PMNs was even higher than freshly isolated ones and was preserved for at least 6 hours after defrosting.

Conclusion

• The outcome of the study supports the potential use of cryopreservation of neutrophils as a viable and effective treatment for mares vulnerable to endometritis.
• The method can potentially offer more efficient therapeutic approaches for endometritis, compared to conventional treatments.