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Hiroyuki Kamao, Atsushi Miki, Junichi Kiryu; Rock inhibitor-induced enhancement of retinal pigment epithelial cell adhesion. Invest. Ophthalmol. Vis. Sci. 2018;59(9):5325.
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© ARVO (1962-2015); The Authors (2016-present)
For the transplantation of human retinal pigment epithelium (RPE), improving cell adhesion is essential, and the focal adhesion (FA) can serve as a good indicator of this status. The aim of this study was to evaluate the effect of Rho kinase (ROCK) inhibitor Y27632 on FA of RPE in vitro.
Human RPE cells were cultured in media with or without Y27632. A luminescent cell viability assay was performed to investigate the effect of Y27632 on cell adhesion. The FA evaluated were the mean size of each cell and FA, number of FA per cell, and total area of FA per cell. These results were evaluated by immunofluorescence images of vinculin, which is one of the markers of FA. The expression levels of vinculin in RPE with or without Y27632 were also studied by real-time RT-PCR.
The number of adhered cell was increased by supplementation of 10 µM Y27632. The mean size of each cell and FA, number of FA per cell, and total area of FA per cell was 1990.8 µm2, 2.48 µm2, 54.0 spots, and 387.0 µm2 in the control, and 2969.6 µm2, 0.77 µm2, 98.4 spots, and 746.7 µm2 in 10 µM Y27632-treated RPE, respectively. Expression levels of vinculin in Y27632-treated RPE were 1.51-fold higher than those in the control.
The inhibition of ROCK signaling by 10 µM Y27632 promoted cell adhesion due to the increase of the number and total area of FA per cell and the expression levels of vinculin. Thus, 10 µM Y27632 could be a great potential supplement for RPE transplantation.
This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.
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