COLONY FORMATION BY HUMAN T-LYMPHOCYTES IN AGAR MEDIUM

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  • 1 January 1977
    • journal article
    • research article
    • Vol. 28  (3) , 526-534
Abstract
An improved method is described for growing human T[thymus-derived]-lymphocyte colonies in agar medium containing phytohemagglutinin (PHA) and sheep red blood cells (SRBC). Cluster and colony growth was obtained when blood mononuclear cells were plated directly in the agar-medium (1-step procedure) or after incubation of cells in liquid medium with PHA (2-step procedure). In the 1-step procedure approximately 1/100 cells plated formed a cluster containing 4-50 cells. In the 2-step procedure 1/20 cells plated formed a cluster or a colony (more than 50 cells). The proliferating cells were sheep-erythrocyte rosette-forming cells (E-RFC). Optimal proliferation was dependent on the presence of phagocytic cells in the cell suspensions cultured. No growth occurred in cultures depleted of E-RFC. Detailed studies of the cycle, velocity sedimentation and density of the cells plated showed that the majority of cluster- and colony-forming cells were small non-cycling lymphocytes with a sedimentation velocity of 4 mm/h, and density between 1.069 and 1.077 g/cm3.