Hypotonically induced changes in the plasma membrane of cultured mammalian cells

Sukhorukov, Vladimir L.; Arnold, W.M.; Zimmermann, U.

doi:10.1007/bf00233049

Cited by 95 publications

(76 citation statements)

References 22 publications

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“…33,40,41,43 In mouse myeloma cells NS1 no swelling and regulatory volume decrease were observed. Instead, NS1 cells started to bleb and express non-spherical shapes ( Figure 3).…”

Section: Cell Viability After Hypotonic Treatmentmentioning

confidence: 94%

“…The increase in cell size requires the unfolding of undulations and invaginations of cell membrane. 42,43 This decrease in membrane undulations results in the decrease of undulation repulsive forces, yielding better cell-cell contacts. 31,[44][45][46][47] Differences among cell lines were found in regulatory volume decrease dynamics.…”

Section: Cell Viability After Hypotonic Treatmentmentioning

confidence: 99%

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Cell size dynamics and viability of cells exposed to hypotonic treatment and electroporation for electrofusion optimization

Ušaj¹,

Trontelj²,

Hudej³

et al. 2009

Radiology and Oncology

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“…33,40,41,43 In mouse myeloma cells NS1 no swelling and regulatory volume decrease were observed. Instead, NS1 cells started to bleb and express non-spherical shapes ( Figure 3).…”

Section: Cell Viability After Hypotonic Treatmentmentioning

confidence: 94%

Section: Cell Viability After Hypotonic Treatmentmentioning

confidence: 99%

Cell size dynamics and viability of cells exposed to hypotonic treatment and electroporation for electrofusion optimization

Ušaj¹,

Trontelj²,

Hudej³

et al. 2009

Radiology and Oncology

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“…The decrease of the mean C, value (and its CV) in hypotonically swollen cells is due to the disappearance of membrane folds and microvilli (12,19,37,38). At physiological osmolality (280-300 mOsm), large variations in the plasma membrane morphology cause the CV of the individual C , values to be higher.…”

Section: Plasma Membrane Capacitymentioning

confidence: 99%

Electropermeabilization and fluorescent tracer exchange: The role of whole‐cell capacitance

et al. 1995

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Transmembrane crossing of charged fluorescent tracers such as propidium iodide (PI) and carboxyfluorescein (CF) can be used to quantitate membrane permeabilization. Murine myeloma Sp2/0-Ag14 cells were loaded with CF (0.1 fmolkell) before electropulsation (0.5-3.0 kV/cm, 40 ps) in medium containing 25-50 pg/ml PI at 21-23°C. Cytograms of PI vs. CF fluorescence showed three readily distinguishable subpopulations: 1) intact living cells with CF but without PI (these form >95% of the prepulsed population), 2) transiently electropermeabilized but resealed cells showing both CF and low-level PI fluorescence, and 3) permanently permeabilized cells without CF but with very high PI fluorescence. Despite the ready influx of PI, the efflux of CF from transiently permeabilized cells was negligible and was insensitive to pulse parameters; however, electrically killed cells (subpopulation 3) lost all CF fluorescence and probably lost their cytoplasm. This difference in transmembrane passage of the dyes is best explained by binding of intracellular CF to macromolecules (and/or organelles). In isotonic "pulse medium," the membranes resealed after electropulsing with a time constant (73 of about 2 min. In 150 mOsm medium, resealing was faster (typically T~ -0.5 min). The population distribution of PI uptake [coefficient of variation (CV) > 40%] was very broad and could not be accounted for by the radius de-10%). The variability in PI uptake could be explained if the electrical energy of the charged membrane, which depends on the whole-cell capacitance (C,), was taken into account. Evaluation of the C, values with single-cell resolution was based on measurement of the electrical charging time constant of the plasma membrane by electrorotation. Key terms: Plasma membrane, electric breakdown, cytoplasm, fluorescent staining, propidium iodide, dye uptake, electrorotation, carboxyfluorescein, flow cytometry, cell viability Electropermeabilization allows the introduction of foreign substances (e.g., drugs, hormones, proteins, plasmids) into living cells (for reviews, see 27,44,45). This technique has been reported to be the most reliable for studying the effects of normally impermeant molecules on the regulation of cell metabolism. Other permeabilization methods (e.g., the use of bacterial toxins, detergents, hypertonic treatment) can markedly affect cell viability and cause undesirable leakage of the cytoplasm or extensive cell lysis ( 2 5 3 0 ) .Plasma membrane electropermeabilization within statistically representative cell samples can be assessed by measuring membrane crossing of tracer molecules in a flow cytometer. Extremely heterogeneous responses to electropulsing have been found in rather homogeneous (origin, size, morphology) cell samples despite the use of uniform electric fields (parallel plate electrodes) for several cell types (6,9,10,24). Flow cytometry of human red blood cells after electropulsation in the presence of FITCdextran (70 kDa) gave rise to at least three distinct subpopulations: ghosts with negligible upta...

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“…The polarizability of living cells depends strongly on their composition, morphology, and phenotype and is also highly dependent on the frequency of the applied electrical field. 26,27 This means that for a given particle type and suspending medium, the particle can experience, at a certain ac frequency applied to the electrodes, a translational force directed toward regions of high electric field strength ͑this phenomenon is called positive DEP, i.e., pDEP͒. Alternatively, by simply changing the frequency, they may experience a force that will direct the particle away from the high electric field strength regions ͑this phenomenon is called negative DEP, i.e., nDEP͒.…”

Section: Introductionmentioning

confidence: 99%

Dielectrophoretic separation of colorectal cancer cells

Yang

Hong

et al. 2010

Biomicrofluidics

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Separation of colorectal cancer cells from other biological materials is important for stool-based diagnosis of colorectal cancer. In this paper, we use conventional dielectrophoresis in a microfluidic chip to manipulate and isolate HCT116 colorectal cancer cells. It is noticed that at a particular alternating current frequency band, the HCT116 cells are clearly deflected to a side channel from the main channel after the electric activation of an electrode pair. This motion caused by negative dielectrophoresis can be used to simply and rapidly separate cancer cells from other cells. In this manuscript, we report the chip design, flow conditions, dielectrophoretic spectrum of the cancer cells, and the enrichment factor of the colorectal cancer cells from other cells.

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Hypotonically induced changes in the plasma membrane of cultured mammalian cells

Cited by 95 publications

References 22 publications

Cell size dynamics and viability of cells exposed to hypotonic treatment and electroporation for electrofusion optimization

Cell size dynamics and viability of cells exposed to hypotonic treatment and electroporation for electrofusion optimization

Electropermeabilization and fluorescent tracer exchange: The role of whole‐cell capacitance

Dielectrophoretic separation of colorectal cancer cells

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