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Appl Environ Microbiol. 1971 May; 21(5): 928-933
Copyright © 1971 American Society for Microbiology. All Rights Reserved.

Growth and Metabolism of L Cells in a Chemically Defined Medium in a Controlled Environment Culture System

I. Effects of O₂ Tension on L-Cell Cultures

Biological Sciences Laboratories, U.S. Army Biological Defense Research Center, Fort Detrick, Frederick, Maryland 21701

ABSTRACT

Six water-jacketed 500-ml Bellco spinner flasks were equipped to monitor and control environmental variables to study their effects on the growth and metabolism of mammalian cells. Studies with automated control of pO₂ levels of L-cell cultures, grown at pH 6.9 ± 0.1, showed that dissolved O₂ tensions of ca. 9% were optimal for cell growth. At pO₂ values of 5 and 20%, maximum cell yields as well as growth rates were reduced by approximately 20%. Peak yields of L-cell cultures exceeded 5 x 10⁶ cells/ml when grown for 4 days without medium renewal from inocula of ca. 10⁶ cells/ml in a defined medium sparged with 5% CO₂ and maintained at 9% dissolved O₂ tension. The redox potentials of L-cell cultures reflected the pO₂ levels in the medium and ranged from –45 to +160 mv (versus calomel reference) for O₂ values ranging from 2 to 20% dissolved oxygen tension. Increased utilization of glucose per cell occurred in the presence of increased pO₂, whereas minimal accumulation of ammonia occurred with a pO₂ value maintained at 9%.