A micromethod for evaluating turkey lymphocyte response to phytomitogens.

A microculture system used in conjunction with a semiautomatic sample harvester is described to determine the in vitro properties of turkey peripheral blood lymphocytes. By this new procedure, multiple tests were done rapidly, relatively few cells were used, and results were highly reproducible. Analysis indicated that the conditions for optimal concanavalin A (con A) stimulation, as measured by incorporation of 3H-thymidine (3H-TdR) included (a) 2 times 10(6) cells per culture in RPMI 1640 medium in the absence of any serum; (b) 0.4 mug of con A per culture, incubated in flat-bottom microtitration wells for 72 hours at 37 C; and (c) 1 muCi of 3H-TdR per culture added 12 to 24 hours before termination. Conditions for optimal stimulation with pokeweed mitogen were similar to those used for con A. The exceptions were that 1 times 10(6) cells per culture were incubated in round-bottom microtitration wells. The pokeweed mitogen gave the highest degree of stimulation when used at a concentration of 80 mug per culture.