Theoretical and Experimental Analysis of Electroporated Membrane Conductance in Cell Suspension

An intense electric field can be applied to increase the membrane conductance G and consequently, the conductivity of cell suspension. This phenomenon is called electroporation. This mechanism is used in a wide range of medical applications, genetic engineering, and therapies. Conductivity measurements of cell suspensions were carried out during application of electric fields from 40 to 165 kV/m. Experimental results were analyzed with two electroporation models: the asymptotic electroporation model was used to estimate Gm at the beginning and at the end of electric field pulse, and the extended Kinosita electroporation model to increase Gm linearly in time. The maximum G was 1-7 × 104 S/m2, and the critical angle (when the Gm is insignificant) was 50°-65°. In addition, the sensitivity of electroporated membrane conductance to extracellular and cytoplasmatic conductivity and cell radius has been studied. This study showed that external conductivity and cell radius are important parameters affecting the pore-opening phenomenon. However, if the cell radius is larger than 7 μm in low conductivity medium, the cell dimensions are not so important.

[1]  L. Chernomordik,et al.  The electrical breakdown of cell and lipid membranes: the similarity of phenomenologies. , 1987, Biochimica et biophysica acta.

[2]  J. Kroeger,et al.  Curvature-driven pore growth in charged membranes during charge-pulse and voltage-clamp experiments. , 2009, Biophysical journal.

[3]  H. Itoh,et al.  Electroporation of cell membrane visualized under a pulsed-laser fluorescence microscope. , 1988, Biophysical journal.

[4]  A. T. Esser,et al.  Membrane electroporation: The absolute rate equation and nanosecond time scale pore creation. , 2006, Physical review. E, Statistical, nonlinear, and soft matter physics.

[5]  Á. Delgado,et al.  Electrokinetics of Concentrated Suspensions of Spherical Colloidal Particles: Effect of a Dynamic Stern Layer on Electrophoresis and DC Conductivity , 2001 .

[6]  D Miklavcic,et al.  The influence of medium conductivity on electropermeabilization and survival of cells in vitro. , 2001, Bioelectrochemistry.

[7]  T. Tsong,et al.  Formation and resealing of pores of controlled sizes in human erythrocyte membrane , 1977, Nature.

[8]  Wanda Krassowska,et al.  Asymptotic model of electroporation , 1999 .

[9]  Airton Ramos.  Effect of the electroporation in the field calculation in biological tissues. , 2005, Artificial organs.

[10]  J. Gimsa,et al.  Dielectric spectroscopy of human erythrocytes: investigations under the influence of nystatin. , 1994, Biophysical journal.

[11]  S Takashima,et al.  Dielectric properties of mouse lymphocytes and erythrocytes. , 1989, Biochimica et biophysica acta.

[12]  I. G. Abidor,et al.  Studies of cell pellets: II. Osmotic properties, electroporation, and related phenomena: membrane interactions. , 1994, Biophysical journal.

[13]  H. Itoh,et al.  Time courses of cell electroporation as revealed by submicrosecond imaging of transmembrane potential. , 1993, Biophysical journal.

[14]  Damijan Miklavčič,et al.  Electric Field-Induced Transmembrane Potential Depends on Cell Density and Organizatio , 1998 .

[15]  D. Miklavčič,et al.  Effective conductivity of cell suspensions , 2002, IEEE Transactions on Biomedical Engineering.

[16]  A. Yaari.  Mobility of human red blood cells of different age groups in an electric field. , 1969, Blood.

[17]  W. Krassowska,et al.  Electrical energy required to form large conducting pores. , 2003, Bioelectrochemistry.

[18]  Mojca Pavlin,et al.  Effective conductivity of a suspension of permeabilized cells: a theoretical analysis. , 2003, Biophysical journal.

[19]  Mojca Pavlin,et al.  Effect of cell electroporation on the conductivity of a cell suspension. , 2005, Biophysical journal.

[20]  J. Weaver,et al.  Transport lattice approach to describing cell electroporation: use of a local asymptotic model , 2004, IEEE Transactions on Plasma Science.

[21]  D. Chang.  2 – Structure and Dynamics of Electric Field-Induced Membrane Pores as Revealed by Rapid-Freezing Electron Microscopy , 1991 .

[22]  Ohshima.  Electrical Conductivity of a Concentrated Suspension of Spherical Colloidal Particles. , 1999, Journal of colloid and interface science.

[23]  Mechanism for the conductivity changes caused by membrane electroporation of CHO cell - pellets , 2004 .

[24]  M. Rols,et al.  Mechanisms of cell membrane electropermeabilization: a minireview of our present (lack of ?) knowledge. , 2005, Biochimica et biophysica acta.

[25]  E. Neumann,et al.  Ionic conductivity of electroporated lipid bilayer membranes. , 2002, Bioelectrochemistry.

[26]  Norde,et al.  Conductivity and Dielectric Dispersion of Gram-Positive Bacterial Cells , 1997, Journal of colloid and interface science.

[27]  Tadej Kotnik,et al.  Sensitivity of transmembrane voltage induced by applied electric fields—A theoretical analysis , 1997 .

[28]  D Krebs,et al.  Electro-rotation of mouse oocytes: single-cell measurements of zona-intact and zona-free cells and of the isolated zona pellucida. , 1987, Biochimica et biophysica acta.

[29]  A. Ramos,et al.  Numerical study of the electrical conductivity and polarization in a suspension of spherical cells. , 2006, Bioelectrochemistry.

[30]  Siewert J Marrink,et al.  Molecular dynamics simulations of hydrophilic pores in lipid bilayers. , 2004, Biophysical journal.

[31]  V. F. Pastushenko,et al.  247 - Electric breakdown of bilayer lipid membranes II. Calculation of the membrane lifetime in the steady-state diffusion approximation , 1979 .

[32]  L. Chernomordik,et al.  Reversible electrical breakdown of lipid bilayers: formation and evolution of pores. , 1988, Biochimica et biophysica acta.

[33]  J Teissié,et al.  An experimental evaluation of the critical potential difference inducing cell membrane electropermeabilization. , 1993, Biophysical journal.

[34]  R P Joshi,et al.  Improved energy model for membrane electroporation in biological cells subjected to electrical pulses. , 2002, Physical review. E, Statistical, nonlinear, and soft matter physics.

[35]  R. Benz,et al.  Kinetics of pore size during irreversible electrical breakdown of lipid bilayer membranes. , 1993, Biophysical journal.

[36]  J. Teissié,et al.  Osmotically induced membrane tension facilitates the triggering of living cell electropermeabilization. , 2004, Bioelectrochemistry.

[37]  A. Ramos,et al.  Numerical simulation of electroporation in spherical cells. , 2004, Artificial organs.

[38]  W. Krassowska,et al.  Modeling electroporation in a single cell. , 2007, Biophysical journal.

[39]  W. Krassowska,et al.  Modeling electroporation in a single cell. I. Effects Of field strength and rest potential. , 1999, Biophysical journal.

[40]  J. Weaver,et al.  Theory of electroporation: A review , 1996 .