Electric impedance of human embryonic stem cell-derived retinal pigment epithelium

The barrier properties of epithelium are conventionally defined by transepithelial resistance (TER). TER provides information about the tightness of the epithelium. Electrical impedance spectroscopy (EIS) provides additional information regarding cell membrane properties, such as changes in electric capacitance and possible parallel or serial pathways that may correlate with the morphology of the cell layer. This study presents EIS of retinal pigment epithelial (RPE) cell model of the putative RPE differentiated from human embryonic stem cells (hESC-RPE). The generally utilized RPE cell model, ARPE-19, was used as immature control. The measured EIS was analyzed by fitting an equivalent electrical circuit model describing the resistive and capacitive properties of the RPE. Our results indicated that TER of hESC-RPE cells was close to the values of human RPE presented in the literature. This provides evidence that the stem cell-derived RPE in vitro can reach high-barrier function. Furthermore, hESC-RPE cells produced impedance spectra that can be modeled by the equivalent circuit of one time constant. ARPE-19 cells produced low-barrier properties, that is, an impedance spectra that suggested poor maturation of ARPE-19 cells. To conclude, EIS could give us means for non-invasively estimating the functionality and maturation of differentiated-RPE cells.

[1]  Joachim Wegener,et al.  Use of electrochemical impedance measurements to monitor β-adrenergic stimulation of bovine aortic endothelial cells , 1999, Pflügers Archiv.

[2]  L. Rizzolo Development and role of tight junctions in the retinal pigment epithelium. , 2007, International review of cytology.

[3]  H. Galla,et al.  Impedance analysis of epithelial and endothelial cell monolayers cultured on gold surfaces. , 1996, Journal of biochemical and biophysical methods.

[4]  Dean Bok,et al.  Na,K-ATPase inhibition alters tight junction structure and permeability in human retinal pigment epithelial cells. , 2003, American journal of physiology. Cell physiology.

[5]  J. Schulzke,et al.  Ussing chamber for high-frequency transmural impedance analysis of epithelial tissues. , 1997, Journal of biochemical and biophysical methods.

[6]  Olaf Strauss,et al.  The retinal pigment epithelium in visual function. , 2005, Physiological reviews.

[7]  H. Tähti,et al.  Evaluation of the selected barrier properties of retinal pigment epithelial cell line ARPE-19 for an in-vitro blood-brain barrier model , 2008, Human & experimental toxicology.

[8]  F. Holz,et al.  Keypathophysiologic pathways in age-related macular disease , 2004, Graefe's Archive for Clinical and Experimental Ophthalmology.

[9]  H. Tähti,et al.  Development of an in vitro blood-brain barrier model-cytotoxicity of mercury and aluminum. , 2004, Toxicology and applied pharmacology.

[10]  S S Miller,et al.  Ion transport mechanisms in native human retinal pigment epithelium. , 1992, Investigative ophthalmology & visual science.

[11]  R. Lund,et al.  Long-term preservation of cortically dependent visual function in RCS rats by transplantation , 2002, Nature Neuroscience.

[12]  R. Lund,et al.  Human embryonic stem cell-derived cells rescue visual function in dystrophic RCS rats. , 2006, Cloning and stem cells.

[13]  B. McKay,et al.  Separation of phenotypically distinct subpopulations of cultured human retinal pigment epithelial cells. , 1994, Experimental cell research.

[14]  Joachim Wegener,et al.  Bioelectrical impedance assay to monitor changes in cell shape during apoptosis. , 2004, Biosensors & bioelectronics.

[15]  S A Lewis,et al.  A spectroscopic method for assessing confluence of epithelial cell cultures. , 1991, The American journal of physiology.

[16]  R. Caldwell,et al.  Retinal pigment epithelial cells from dystrophic rats form normal tight junctions in vitro. , 1997, Investigative ophthalmology & visual science.

[17]  E. Schifferdecker,et al.  The AC impedance of necturus gallbladder epithelium , 1978, Pflügers Archiv.

[18]  E. Barrón,et al.  A protocol for the culture and differentiation of highly polarized human retinal pigment epithelial cells , 2009, Nature Protocols.

[19]  F. Chen,et al.  RPE transplantation and its role in retinal disease , 2007, Progress in Retinal and Eye Research.

[20]  A. Ahmad,et al.  Membrane polarity of the Na(+)-K+ pump in primary cultures of Xenopus retinal pigment epithelium. , 1994, Experimental eye research.

[21]  Michael Fromm,et al.  Two-path impedance spectroscopy for measuring paracellular and transcellular epithelial resistance. , 2009, Biophysical journal.

[22]  L. Sherwood Human Physiology : From Cells to Systems , 1989 .

[23]  H. Skottman Derivation and characterization of three new human embryonic stem cell lines in Finland , 2010, In Vitro Cellular & Developmental Biology - Animal.

[24]  D P Joseph,et al.  Apical and basal membrane ion transport mechanisms in bovine retinal pigment epithelium. , 1991, The Journal of physiology.

[25]  H. Uusitalo,et al.  Toward the defined and xeno-free differentiation of functional human pluripotent stem cell–derived retinal pigment epithelial cells , 2011, Molecular vision.

[26]  A. Urtti,et al.  Filter-cultured ARPE-19 cells as outer blood-retinal barrier model. , 2010, European journal of pharmaceutical sciences : official journal of the European Federation for Pharmaceutical Sciences.

[27]  Arvydas Maminishkis,et al.  Confluent monolayers of cultured human fetal retinal pigment epithelium exhibit morphology and physiology of native tissue. , 2006, Investigative ophthalmology & visual science.

[28]  C. Hsieh,et al.  Polarized secretion of PEDF from human embryonic stem cell-derived RPE promotes retinal progenitor cell survival. , 2011, Investigative ophthalmology & visual science.

[29]  Joachim Wegener,et al.  Barrier function of porcine choroid plexus epithelial cells is modulated by cAMP-dependent pathways in vitro , 2000, Brain Research.

[30]  D. Frambach,et al.  Beta Adrenergic Receptors on Cultured Humon Retinal Pigment Epithelium , 2005 .

[31]  L. Hjelmeland,et al.  ARPE-19, a human retinal pigment epithelial cell line with differentiated properties. , 1996, Experimental eye research.