A technique for collection and cultivation of macrophages from Cynomolgus monkeys (Macaca fascicularis).

Several techniques were compared for collection of peritoneal mononuclear phagocytes from cynomolgus monkeys. A multiple-holed cannula proved superior to a hypodermic needle for the collection of peritoneal washings because it was not occluded by the omentum. With the cannula technique 55--70% of the harvest medium could be recovered from the peritoneal cavity with a yield of 2.5--5 x 10(5) macrophages per ml. A closed system for injecting and collecting harvest medium was better than a syringe in preventing bacterial contamination of peritoneal washings. After collection, cells were resuspended in growth medium to a concentration of 5 x 10(5) macrophages per ml, distributed into Leighton tubes, and incubated at 36 degrees C. After 12 hours, cultures were washed to remove non-adherent cells and refed with growth medium. This technique provided a sterile, reliable method for the collection and cultivation of macrophages from monkeys.