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Resuspension-induced ion flux
1New Ideas, Co., Rockville, Maryland 20852, USA.
Analytical Biochemistry
|February 19, 1998
Summary
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.
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.
- 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.
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