Quantitative Model for Ion Transport and Cytoplasm Conductivity of Chinese Hamster Ovary Cells

Overview

Journal Sci Rep

Specialty Science

Date 2018 Dec 15

PMID 30546044

Citations 3

Authors

Azita Fazelkhah

Katrin Braasch

Samaneh Afshar

Elham Salimi

Michael Butler

Greg Bridges

Douglas Thomson

Affiliations

Soon will be listed here.

Abstract

In mammalian cells cytoplasm ion concentrations and hence cytoplasm conductivity is an important indicator of their physiological state. Changes in the cytoplasm conductivity has been associated with physiological changes such as progression of cancer and apoptosis. In this work, a model that predicts the effects of physiological changes in ion transport on the cytoplasm conductivity of Chinese hamster ovary (CHO) cells is demonstrated. We determined CHO-specific model parameters, Na/K ATPase pumps and ion channels densities, using a flux assay approach. The obtained sodium (P), potassium (P) and chloride (P) permeability and Na/K ATPase pump density were estimated to be 5.6 × 10 cm/s, 5.6 × 10 cm/s, 3.2 × 10 cm/s and 2.56 × 10 mol/cm, respectively. The model was tested by comparing the model predictions with the experimentally determined temporal changes in the cytoplasm conductivity of Na/K ATPase pump inhibited CHO cells. Cells' Na/K ATPase pumps were inhibited using 5 mM Ouabain and the temporal behavior of their cytoplasm conductivity was measured using dielectrophoresis cytometry. The measured results are in close agreement with the model-calculated values. This model will provide insight on the effects of processes such as apoptosis or external media ion concentration on the cytoplasm conductivity of mammalian cells.

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References

Jakobsson E . Interactions of cell volume, membrane potential, and membrane transport parameters. Am J Physiol. 1980; 238(5):C196-206. DOI: 10.1152/ajpcell.1980.238.5.C196. View

Coley H, Labeed F, Thomas H, Hughes M . Biophysical characterization of MDR breast cancer cell lines reveals the cytoplasm is critical in determining drug sensitivity. Biochim Biophys Acta. 2007; 1770(4):601-8. DOI: 10.1016/j.bbagen.2006.12.002. View

Zhou Y, Basu S, Laue E, Seshia A . Single cell studies of mouse embryonic stem cell (mESC) differentiation by electrical impedance measurements in a microfluidic device. Biosens Bioelectron. 2016; 81:249-258. PMC: 4833703. DOI: 10.1016/j.bios.2016.02.069. View

Mulhall H, Cardnell A, Hoettges K, Labeed F, Hughes M . Apoptosis progression studied using parallel dielectrophoresis electrophysiological analysis and flow cytometry. Integr Biol (Camb). 2015; 7(11):1396-401. DOI: 10.1039/c5ib00109a. View

Orlov S, Platonova A, Hamet P, Grygorczyk R . Cell volume and monovalent ion transporters: their role in cell death machinery triggering and progression. Am J Physiol Cell Physiol. 2013; 305(4):C361-72. DOI: 10.1152/ajpcell.00040.2013. View

Obergrussberger A, Stolzle-Feix S, Becker N, Bruggemann A, Fertig N, Moller C . Novel screening techniques for ion channel targeting drugs. Channels (Austin). 2015; 9(6):367-75. PMC: 4850050. DOI: 10.1080/19336950.2015.1079675. View

Fraser J, Huang C . Quantitative techniques for steady-state calculation and dynamic integrated modelling of membrane potential and intracellular ion concentrations. Prog Biophys Mol Biol. 2006; 94(3):336-72. DOI: 10.1016/j.pbiomolbio.2006.10.001. View

Gardette R, Debono M, Dupont J, Crepel F . Electrophysiological studies on the postnatal development of intracerebellar nuclei neurons in rat cerebellar slices maintained in vitro. II. Membrane conductances. Brain Res. 1985; 352(1):97-106. DOI: 10.1016/0165-3806(85)90091-4. View

Farinas J, Kneen M, Moore M, Verkman A . Plasma membrane water permeability of cultured cells and epithelia measured by light microscopy with spatial filtering. J Gen Physiol. 1997; 110(3):283-96. PMC: 2229369. DOI: 10.1085/jgp.110.3.283. View

10.

Moller B, Vaag A, Johansen T . Ouabain inhibition of the sodium-potassium pump: estimation of ED50 in different types of human leucocytes in vitro. Br J Clin Pharmacol. 1990; 29(1):93-100. PMC: 1380066. DOI: 10.1111/j.1365-2125.1990.tb03607.x. View

11.

Salimi E, Braasch K, Butler M, Thomson D, Bridges G . Dielectric model for Chinese hamster ovary cells obtained by dielectrophoresis cytometry. Biomicrofluidics. 2016; 10(1):014111. PMC: 4723405. DOI: 10.1063/1.4940432. View

12.

Terashima K, Takeuchi A, Sarai N, Matsuoka S, Shim E, Leem C . Modelling Cl- homeostasis and volume regulation of the cardiac cell. Philos Trans A Math Phys Eng Sci. 2006; 364(1842):1245-65. DOI: 10.1098/rsta.2006.1767. View

13.

Henslee E, Torcal Serrano R, Labeed F, Jabr R, Fry C, Hughes M . Accurate quantification of apoptosis progression and toxicity using a dielectrophoretic approach. Analyst. 2016; 141(23):6408-6415. DOI: 10.1039/c6an01596d. View

14.

Gamper N, Stockand J, Shapiro M . The use of Chinese hamster ovary (CHO) cells in the study of ion channels. J Pharmacol Toxicol Methods. 2005; 51(3):177-85. DOI: 10.1016/j.vascn.2004.08.008. View

15.

Altomare L, Borgatti M, Medoro G, Manaresi N, Tartagni M, Guerrieri R . Levitation and movement of human tumor cells using a printed circuit board device based on software-controlled dielectrophoresis. Biotechnol Bioeng. 2003; 82(4):474-9. DOI: 10.1002/bit.10590. View

16.

Salimi E, Braasch K, Fazelkhah A, Afshar S, Saboktakin Rizi B, Mohammad K . Single cell dielectrophoresis study of apoptosis progression induced by controlled starvation. Bioelectrochemistry. 2018; 124:73-79. DOI: 10.1016/j.bioelechem.2018.07.003. View

17.

Xu J, Wang X, Ensign B, Li M, Wu L, Guia A . Ion-channel assay technologies: quo vadis?. Drug Discov Today. 2001; 6(24):1278-1287. DOI: 10.1016/s1359-6446(01)02095-5. View

18.

Duncan L, Shelmerdine H, Hughes M, Coley H, Hubner Y, Labeed F . Dielectrophoretic analysis of changes in cytoplasmic ion levels due to ion channel blocker action reveals underlying differences between drug-sensitive and multidrug-resistant leukaemic cells. Phys Med Biol. 2008; 53(2):N1-7. DOI: 10.1088/0031-9155/53/2/N01. View

19.

Armstrong C . The Na/K pump, Cl ion, and osmotic stabilization of cells. Proc Natl Acad Sci U S A. 2003; 100(10):6257-62. PMC: 156359. DOI: 10.1073/pnas.0931278100. View

20.

Opel C, Li J, Amanullah A . Quantitative modeling of viable cell density, cell size, intracellular conductivity, and membrane capacitance in batch and fed-batch CHO processes using dielectric spectroscopy. Biotechnol Prog. 2010; 26(4):1187-99. DOI: 10.1002/btpr.425. View