Control of cellular proliferation in HeLa-S3 suspension cultures. Characterization of cultures utilizing acridine orange staining procedures

Journal of Cellular Physiology
K D Bauer, L A Dethlefsen

Abstract

Growth control is investigated in detail in fed and unfed HeLa-S3 suspension cultures. Two-step acridine orange staining and flow cytometric analysis indicated 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 Gi-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 345 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...Continue Reading

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Citations

Jan 1, 1983·Pharmacology & Therapeutics·J W Gray
Oct 16, 1999·Journal of Microbiological Methods·R BarthelsonA Mobasseri
May 1, 1988·Cell and Tissue Kinetics·Z DarzynkiewiczK Shogen
Jan 1, 1984·Cell and Tissue Kinetics·C A WallenL A Dethlefsen
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Jan 5, 2002·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·Stanislaw M MikulskiKuslima Shogen
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Sep 1, 1987·Experimental Cell Research·Z DarzynkiewiczP J Higgins

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