Does gradient isolation preserve equine neutrophil viability and oxidative function for up to six hours?

This study investigated isolation of equine neutrophils and the assessment of their antioxidant function using the nitroblue tetrazolium (NBT) reduction test, an indicator of reactive oxygen species (ROS) activation. The objective was to test the viability of neutrophils isolated from equine whole blood and their ability to undergo respiratory burst, comparing the results with whole blood. The hypothesis tested was that isolation of equine neutrophils allows for effective functional evaluation, even after isolation process, with no significant differences between whole blood and isolated cells. The study showed that neutrophils maintained their ability to reduce NBT dye over a 6-hour period, with no significant differences between time points analyzed (0 h, 3 h, and 6 h). Isolation was performed using Histopaque®, and total neutrophil concentration and oxidative function were evaluated at three different time points. Cell counts and viability assessments were conducted using both automated and manual techniques, confirming the prolonged viability of neutrophils and the accuracy of antioxidant function testing.