Control of cellular proliferation in HeLa‐S3 suspension cultures. Characterization of cultures utilizing acridine orange staining procedures
- 1 July 1981
- journal article
- research article
- Published by Wiley in Journal of Cellular Physiology
- Vol. 108 (1), 99-112
- https://doi.org/10.1002/jcp.1041080113
Abstract
Growth control is investigated in detail in fed and unfed HeLa‐S3 suspension cultures. Two‐step acridine orange staining and flow cytometric analysis indicate declines in cellular red fluorescence (proportional to RNA content) of 40–50% between exponential and plateau phase in both culture types. Cellular green fluorescence (DNA content) assessed simultaneously indicates an increment of cells with G1‐DNA content in plateau phase in the unfed cultures, while fed cultures show a brief increment in G1‐phase cells in the transition phase followed by a recovery in plateau phase to a value similar to that of exponential cultures. Temporal declines in the 3H‐thymidine pulse‐labeling index are observed in both culture systems. These data along with the flow cytometry data indicate a distinct G1‐arrest in the unfed plateau cultures and suggest a random arrest of cells about the cell cycle in fed plateau cultures. Acidic acridine orange staining and flow cytometric analysis furthermore indicate the occurrence of a quiescent population comprising approximately 34% of the total cells and consisting of both dead and viable cells in plateau phase unfed cultures. In contrast, fed plateau cultures show approximately 14% quiescent, mostly dead cells. Also, both culture systems show temporal declines in the clonogenic index and a longer cell‐cycle transit time in plateau phase relative to exponential phase. These findings confirm earlier work which indicates that the environment has a profound influence on the mode of growth control for mammalian cells in vitro.This publication has 25 references indexed in Scilit:
- Different sensitivity of chromatin to acid denaturation in quiescent and cycling cells as revealed by flow cytometry.Journal of Histochemistry & Cytochemistry, 1979
- Mapping G1 phase by the structural morphology of the prematurely condensed chromosomesJournal of Cellular Physiology, 1978
- Recognition of cells in mitosis by flow cytofluormetry.Journal of Histochemistry & Cytochemistry, 1977
- Coordinate regulation of protein synthesis and messenger RNA content during growth arrest of suspension Chinese hamster ovary cellsJournal of Cellular Physiology, 1977
- The in vivo reproductive potential of density separated cellsExperimental Cell Research, 1974
- REGULATION OF INITIATION OF DNA SYNTHESIS IN CHINESE HAMSTER CELLSThe Journal of cell biology, 1970
- Fluctuations of DNA-dependent RNA polymerase and synthesis of macromolecules during the growth cycle of Novikoff rat hepatoma cells in suspension cultureJournal of Cellular Physiology, 1969
- Chinese hamster cell monolayer cultures: I. Changes in cell dynamics and modifications of the cell cycle with the period of growthExperimental Cell Research, 1968
- STATIONARY PHASE OF CULTURED MAMMALIAN CELLS (L5178Y)The Journal of cell biology, 1967
- Systematic fluctuations in the cellular protein, RNA and DNA during growth of mammalian cell culturesBiochimica et Biophysica Acta, 1959