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pH changes in pulsed CO2 incubators cause periodic changes in cell morphology

C M Lo1, C R Keese, I Giaever

  • 1Department of Physics and Biology, School of Science, Rensselaer Polytechnic Institute, Troy, New York 12180-3590.

Experimental Cell Research
|August 1, 1994
PubMed
Summary

Small CO2 fluctuations in incubators cause subtle, yet detectable, cell morphology changes. These oscillations impact cell behavior, especially in spent media, affecting paracellular resistance in cultured cells.

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Area of Science:

  • Cell Biology
  • Biophysics

Background:

  • Animal cell culture commonly uses bicarbonate-buffered media with elevated CO2.
  • Incubators maintain CO2 via feedback, causing minor oscillations (approx. 0.5%) in CO2 levels.
  • These oscillations lead to small, previously undetected pH fluctuations in the culture medium.

Purpose of the Study:

  • To investigate the impact of incubator-induced CO2 and pH oscillations on cell morphology and behavior.
  • To determine if subtle cellular changes can be detected and quantified.

Main Methods:

  • Electric Cell-Substrate Impedance Sensing (ECIS) was employed to detect subtle alterations in cell morphology.
  • Fast Fourier Transform (FFT) analysis was used to identify periodicities in cellular changes.
  • A model of cell-electrode interactions was utilized to analyze ECIS measurements.

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Main Results:

  • Periodic cell morphology changes were detected, synchronized with CO2 addition cycles.
  • These effects were more pronounced in partially spent media and with high surface-to-volume ratios.
  • Confluent fibroblasts and endothelial cells showed morphological changes primarily as alterations in paracellular resistance.

Conclusions:

  • Periodic cell morphology changes are influenced by medium condition and vessel geometry.
  • Incubator CO2 oscillations demonstrably affect cell behavior, particularly paracellular resistance in specific cell types.
  • ECIS is a sensitive method for detecting subtle, oscillation-induced cellular responses.