Open AccessA comparative kinetic analysis of proliferation in vitro of Con‐A‐treated splenocytes and syngeneic leukaemia cells
Author(s) -
Olivotto M.,
Boddi V.,
Sbarba P. Dello,
Arcangeli Annarosa
Publication year - 1982
Publication title -
cell proliferation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.647
H-Index - 74
eISSN - 1365-2184
pISSN - 0960-7722
DOI - 10.1111/j.1365-2184.1982.tb01068.x
Subject(s) - thymidine , splenocyte , labelling , in vitro , biology , kinetics , microbiology and biotechnology , population , cell growth , exponential growth , immunology , chemistry , biochemistry , medicine , mathematics , environmental health , quantum mechanics , mathematical analysis , physics
. The growth kinetics of Con‐A‐treated mouse splenocytes and syngeneic leukaemia cells cultured in vitro were compared with respect to (i) the total cell number, (ii) the rate of [ 14 C]thymidine incorporation (measured by pulse‐labelling the cells at various times of incubation), and (iii) the labelling index of the cell populations. By correlating the thymidine incorporation, labelling index and cell number data, it has been established that, for both types of cells, the rate of [ 14 C]thymidine incorporation is directly proportional to the number of cells synthesizing DNA. A new approach to cytokinetic analysis has been developed, showing that important information can be obtained by determining the cumulative kinetics of [ 14 C]thymidine incorporation. The latter has been calculated by integrating the area underneath the time course of the rate of thymidine incorporation, and was directly proportional to the overall growth of both leukaemia cells and Con‐A‐stimulated splenocytes. Based on this proportionality, an estimate of the average duration of the S phase for both types of cells was calculated, suggesting that normal and neoplastic blasts maintain this parameter at a constant value (7.6 and 5.9 hr, respectively) throughout different stages of growth. The percentage of Con‐A‐responsive cells within the initial splenocyte population and their overall proliferation in vitro have been determined by a procedure which measures the cumulative kinetics of thymidine incorporation and the kinetics of cell total number in the presence or in the absence of the lectin, as well as in the presence of Con‐A plus colcemid. A minor fraction (11%) of the initial splenocytes is recruited into cycle by Con‐A, proliferating with similar kinetics to that of leukaemia cells in the same conditions. The great majority of the initial splenocyte population is unaffected by Con‐A, decaying exponentially throughout the incubation with the same half‐life (28 hr), both in the presence or in the absence of the lectin.