In vitro growth of colonies from human peripheral blood lymphocytes stimulated by phytohemagglutinin.

Normal peripheral human white blood cels (WBC) plated in agar culture generate colonies of blastoid cells when stimulated in vitro by phytohemagglutinin (PHA). Aggregates of PHA transformed cells are first visible by day 3-4. By day 7, the morphology of colonies becomes typical as clear discoid raspberry-like aggregates of 50-200 cells or more growing at or near the surface of the agar plate. Their dense arrangement and the coexistence within individual colonies of large and smaller cells are two main characterisitcs of these colonies. Wright-Giemsa stained preparations from cytocentrifuged colonies show cells at different stages of "lymphocyte transformation". These cells are PAS and peroxidase negative. They do not bear surface immunoglobulins but most of them form rosettes with sheep red blood cells. These observations strongly suggest that these colonies are made of T-lymphocytes. However, in view of the number of isolated transformed cells which do not form colonies, the possibility is raised that the cells which generate these colonies may represent a subset of PHA sensitive cells.