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Resuspension-induced ion flux

D B Millar1

  • 1New Ideas, Co., Rockville, Maryland 20852, USA.

Analytical Biochemistry
|February 19, 1998
PubMed
Summary
This summary is machine-generated.

Pelleting and resuspension of Fura 2-labeled cells causes a rapid calcium (Ca) flux, known as resuspension-induced ion flux (RIIF). This calcium influx requires specific cellular conditions and is influenced by pelleting parameters.

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Area of Science:

  • Cellular Biology
  • Biophysics
  • Neuroscience

Background:

  • Fura 2 is a fluorescent indicator used to measure intracellular calcium levels.
  • Glioma cells are a type of tumor cell originating in the brain.
  • Calcium ions play crucial roles in various cellular processes.

Purpose of the Study:

  • To characterize the resuspension-induced ion flux (RIIF) in Fura 2-labeled c6 glioma cells.
  • To identify the conditions and cellular components necessary for the RIIF effect.
  • To investigate the parameters influencing the magnitude of RIIF.

Main Methods:

  • Labeling c6 glioma cells with Fura 2.
  • Inducing ion flux through pelleting and resuspension.
  • Measuring intracellular calcium levels using fluorescence.

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  • Investigating the effect of temperature, external calcium, and channel blockers.

    • Main Results:

    • Pelleting and resuspension induced a significant, transient calcium flux (RIIF).
    • RIIF was dependent on temperature, external calcium, cytoskeletal integrity, and functional ion channels (calcium and potassium).
    • Pelleting speed, indicating cell contact and reduced medium, influenced RIIF magnitude.
    • The RIIF effect was observed in multiple cell types.

    Conclusions:

    • Resuspension-induced ion flux is a reproducible phenomenon in c6 glioma cells.
    • RIIF is a calcium-dependent process requiring specific cellular conditions and functional ion channels.
    • Cellular interactions and medium reduction during pelleting are key factors in RIIF.