Density-dependent effects of oxygen on the growth of mammalian fibroblasts in culture

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Abstract
Various concentrations of oxygen were used to determine the optimum culture medium P for survival and proliferation of attached human and mouse fibroblasts grown from different inoculum sizes. When T−15 flasks were seeded with ⪕ 2 × 104 cells (⪕ 1.3 × 103 cells/cm2), the highest plating efficiencies and cell yields were obtained with a culture medium P of 40—60 mm Hg. At higher inoculum sizes (105 cells per T‐15) used routinely for mass cultures, no difference in cell yield or glycolytic activity was observed between cultures gassed with atmospheric, i.e., 18% O2 (growth medium P ≅ 125—135 mm Hg) and those gassed with 1% O2 (growth medium P ≅ 40—60 mm Hg). The enhanced clonal growth observed at the latter P results from an increased proliferation rate rather than more efficient attachment and survival of inoculated cells. Glucose uptake and lactic acid accumulation were increased in sparse cultures sparged with 1% O2. A slight extension of lifespan was observed in WI−38 cells serially subcultured with a gas phase of 1% O2.