Mononuclear Phagocyte Colony Formation of Human Spleen Cells in Liquid Culture

Abstract

We found that mononuclear phagocytes formed a distinct number of clusters and colonies on the bottom of a culture dish 7 days later but granulocytes did not, when a large number of human spleen cells were cultured in liquid medium. In all gastric cancer bearers and patients with portal hypertension operated on, however, colony formation was restricted to spleen cells from patients with advanced gastric cancer and from a group of patients with portal hypertension. These spleen cells formed mononuclear phagocyte colonies without the help of exogeneous colony stimulating factor (CSF). We further demonstrated that the colony-forming cells were glass non-adherent and nylon wool adherent, and that spontaneous colony formation required cooperation between the colony-forming cells and colony-stimulating cells adherent to a plastic surface.There have been many studies on granulocyte-monocyte (GM) colony formation of murine bone marrow cells since the reports of Bradely and Metcalf (1) and Pluznik and Sachs (23) in 1966. Senn and his colleagues (25) and Pike and Robinson (22) reported GM colony formation of human bone marrow cells in 1967 and 1970, respectively. Although they used a semi-solid agar technique, Stewart and his colleagues reported macrophage colony formation of murine peritoneal exudate cells in a liquid culture in 1975 (26). Since then, some reports using the liquid culture technique have been published (2,8,9,30). This simplified technique is available for detailed analysis of mononuclear phagocyte colony formation. We know little about the aspects of the mononuclear phagocyte system in human spleen. We therefore investigated human splenic mononuclear phagocyte colony formation in liquid culture in this study. MATERIALS AND METHODSPreparation of spleen cell suspensions and cultures. The spleens surgically resected from patients with gastric cancer or with portal hypertension were minced in Eagle's minimum essential medium (MEM) and compressed gently with a rubber police-72 5