Optimization of the under-agarose assay of porcine neutrophil migration

Elliott, G.S.; Rebar, A.H.; McCabe, G.P.; Alzola, C.

Optimization of the under-agarose assay of porcine neutrophil migration

1990

Elliott, G.S. | Rebar, A.H. | McCabe, G.P. | Alzola, C.

Important procedural factors in the under-agarose assay for porcine neutrophil migration were identified, and optimal conditions were established. Three factors were tested: the concentration of zymosan-activated serum inoculated into the outer well; the number of neutrophils inoculated into the center well; and the time of incubation of the agarose plates. All factors had a significant (P < 0.0001, 0.0001, and 0.01, respectively) effect on the chemotactic index of porcine neutrophils. The optimal combination of these 3 factors was undiluted zymosan-activated serum as the chemoattractant, 8 x 10(5) neutrophils inoculated into the center well, and 5 hours of incubation. The assay was validated, using standard conditions, and the data were used to predict the number of pigs and/or repetitive assays needed to identify differences among experimental groups.

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