SEQUENTIAL PROLIFERATION INDUCED IN HUMAN PERIPHERAL-BLOOD LYMPHOCYTES BY MITOGEN .1. GROWTH OF 1000 LYMPHOCYTES IN FEEDER LAYER CULTURES

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  • 1 January 1977
    • journal article
    • research article
    • Vol. 118  (5) , 1763-1767
Abstract
A culture system is described in which 1000 human peripheral blood lymphocytes diluted in 2.5 .times. 105 mitomycin-treated autologous cells respond to phytohemagglutinin (PHA). Proliferation data, including 3H-TdR [thymidine] uptake, cell survival counts, and mitotic indices, indicate that this inoculum expands from 1000 to 40,000 cells by day 7, suggesting 5 or 6 sequential cell divisions. Chromosome markers in allogeneic cultures demonstrate that the dividing cells are derived from the original 1000 cells and not from the feeder layer of mitomycin-treated lymphocytes. The time course of proliferation in this system is similar to that in other human lymphocyte culture systems with a low percentage of responding cells, as in the response to PHA of cells from patients with chronic lymphocytic leukemia or the response for normal lymphocytes to antigens. The conditions provided by the feeder layer which permit proliferation of this small number of lymphocytes are not precisely known, but erythrocytes, heat-killed lymphocytes or inert particles do not provide a satisfactory substitute.