The Journal of Cell Biology, Vol 85, 459-465, Copyright © 1980 by The Rockefeller University Press
Lymphocyte cell-cycle analysis by flow cytometry. Evidence for a specific postmitotic phase before return to G0
DP Richman
We studied the cell cycle of lectin-stimulated human lymphocytes, making
use of a flow cytometer. The RNA and DNA content of large numbers of
individual cells was determined by supravital staining with acridine
orange. The present study confirmed previous observations by others of a
progression from G0 through G1 and S phase to G2/mitosis during the first 3
d in culture. It was also found that on subsequent days stimulated cells,
before their return to G0, remained stationary in a state in which they
contained the G0 complement of DNA and approximately twice the G0
complement of RNA. Cell-cycle manipulation with vinblastine and
5-bromo-2-deoxyuridine (BUdR) revealed that previous passage through both S
phase and mitosis was required for entry into this newly observed late
phase. In addition, there was high correlation (r = 0.973, P less than
0.001) between the number of cells in the late phase and measured
[3H]thymidine uptake. It therefore appears that, in this system, stimulated
cells remain in a distinct cell-cycle phase for a number of hours before
their return to the resting state.
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