Confocal microscopy and electrophysiological study of single patient corneal endothelium cell cultures

Francesca Tatini; Francesca Rossi; Elisabetta Coppi; Giada Magni; Irene Fusco; Luca Menabuoni; Felicita Pedata; Anna Maria Pugliese; Roberto Pini

doi:10.1117/12.2212636

6 April 2016 Confocal microscopy and electrophysiological study of single patient corneal endothelium cell cultures

Francesca Tatini, Francesca Rossi, Elisabetta Coppi, Giada Magni, Irene Fusco, Luca Menabuoni, Felicita Pedata, Anna Maria Pugliese, Roberto Pini

Author Affiliations +

Proceedings Volume 9711, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues IX; 97110G (2016) https://doi.org/10.1117/12.2212636
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

The characterization of the ion channels in corneal endothelial cells and the elucidation of their involvement in corneal pathologies would lead to the identification of new molecular target for pharmacological treatments and to the clarification of corneal physiology. The corneal endothelium is an amitotic cell monolayer with a major role in preserving corneal transparency and in regulating the water and solute flux across the posterior surface of the cornea. Although endothelial cells are non-excitable, they express a range of ion channels, such as voltage-dependent Na⁺ channels and K⁺ channels, L-type Ca² channels and many others. Interestingly, purinergic receptors have been linked to a variety of conditions within the eye but their presence in the endothelium and their role in its pathophysiology is still uncertain. In this study, we were able to extract endothelial cells from single human corneas, thus obtaining primary cultures that represent the peculiarity of each donor. Corneas were from tissues not suitable for transplant in patients. We characterized the endothelial cells by confocal microscopy, both within the intact cornea and in the primary endothelial cells cultures. We also studied the functional role of the purinergic system (adenosine, ATP and their receptors) by means of electrophysiological recordings. The experiments were performed by patch clamp recordings and confocal time-lapse microscopy and our results indicate that the application of purinergic compounds modulates the amplitude of outward currents in the isolated endothelial cells. These findings may lead to the proposal of new therapies for endothelium-related corneal diseases.

Conference Presentation

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Francesca Tatini, Francesca Rossi, Elisabetta Coppi, Giada Magni, Irene Fusco, Luca Menabuoni, Felicita Pedata, Anna Maria Pugliese, and Roberto Pini "Confocal microscopy and electrophysiological study of single patient corneal endothelium cell cultures", Proc. SPIE 9711, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues IX, 97110G (6 April 2016); https://doi.org/10.1117/12.2212636

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