Development of Cell Analysis Software for Cultivated Corneal Endothelial Cells

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To develop analysis software for cultured human corneal endothelial cells (HCECs).


Software was designed to recognize cell borders and to provide parameters such as cell density, coefficient of variation, and polygonality of cultured HCECs based on phase contrast images. Cultured HCECs with high or low cell density were incubated with Ca2+-free and Mg2+-free phosphate-buffered saline for 10 minutes to reveal the cell borders and were then analyzed with software (n = 50).


Phase contrast images showed that cell borders were not distinctly outlined, but these borders became more distinctly outlined after phosphate-buffered saline treatment and were recognized by cell analysis software. The cell density value provided by software was similar to that obtained using manual cell counting by an experienced researcher. Morphometric parameters, such as the coefficient of variation and polygonality, were also produced by software, and these values were significantly correlated with cell density (Pearson correlation coefficients −0.62 and 0.63, respectively).


The software described here provides morphometric information from phase contrast images, and it enables subjective and noninvasive quality assessment for tissue engineering therapy of the corneal endothelium.

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