Chicken Lymphocyte Stimulation by Mitogens: A Microassay with Whole-Blood Cultures

A micromethod is described that uses whole blood for studying mitogen stimulation of chicken lymphocytes. Procedures critical for obtaining a stimulatory response up to 200 times that of unstimulated control cultures include: 1) use of no more than 10 microliter of whole blood per culture; 2) incubation at 41 C for 72 hours in 0.2 ml of medium RPMI 1640 containing 100 microgram of stock phytohemagglutinin-P and 3) pulse labeling for 8 hours with 5-[125I] iodo-2'-deoxyruidine. The method is easy, rapid, and suitable for sequential studies of pathogenesis of disease in poultry.