Purchase this article with an account.
Virpi Savolainen, Kati Juuti-Uusitalo, Niina Onnela, Hanna Vaajasaari, Susanna Narkilahti, Riitta Suuronen, Heli Skottman, Jari Hyttinen; Impedance Spectroscopy for Determining Human Embryonic Stem Cell-Derived Retinal Pigment Epithelium Maturity. Invest. Ophthalmol. Vis. Sci. 2011;52(14):402.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
The epithelial morphogenesis in the development of impermeable retinal pigment epithelium (RPE) monolayer with highly organized tight junctions (TJ) has been shown to be a gradual process that is triggered by the cell-to-cell contact at confluence. The objective of this study was to evaluate the barrier function development in human embryonic stem cell (hESC)-derived retinal pigment epithelial cell cultures.
The barrier function was assessed from hESC-derived RPE cells cultured for different periods of time. The chosen cultures were subjected to the kinetic assessment of barrier properties after calcium sequestration with EGTA. The development of the epithelial was assessed by electric impedance spectroscopy (EIS), fluorescent particle based permeability measurement and microscopic inspection.
The impedance values negatively correlated to the permeability of small molecular weight particles: The most mature cultures had the highest impedance values and lowest permeability; respectively the most immature cultures had the lowest impedance and highest permeability values. Furthermore more mature cells were more persistent against the chemical insult. The EIS of samples of high integrity fitted well to a single equivalent RC circuit model. Also, after EGTA treatment these cells maintained their EIS shape, while EGTA treatment of cell with lower integrity changed EIS shapes indicating opening of secondary pathway for electric currents and possibly particle permeability as their TJs were deteriorating.
In this study we have shown that EIS values correlate well with the cell morphology and permeability, suggesting that EIS can be used to determine the overall integrity, maturity and functionality of cell cultures. This offers a non-invasive way to evaluate the basic functionality of the cell layer which is well warranted in future transplantation therapies and in in vitro cell culture models in drug testing and development.
This PDF is available to Subscribers Only