Development of Cell Analysis Software for Cultivated Corneal Endothelial Cells

Area of Science:

• Ophthalmology
• Biomedical Engineering
• Cell Biology

Background:

• Cultured human corneal endothelial cells (HCECs) are crucial for corneal endothelium tissue engineering.
• Accurate morphometric analysis of HCECs is essential for quality control.
• Existing methods for HCEC analysis can be subjective and time-consuming.

Purpose of the Study:

• To develop and validate analysis software for cultured human corneal endothelial cells (HCECs).
• To enable objective, non-invasive quality assessment of HCECs for tissue engineering applications.

Main Methods:

• Software designed to identify cell borders and quantify parameters like cell density, coefficient of variation, and polygonality from phase contrast images.
• HCECs were treated with Ca-free and Mg-free phosphate-buffered saline to enhance cell border visibility.
• Analysis of 50 cultured HCEC samples using the developed software.

Main Results:

• Phosphate-buffered saline treatment significantly improved the distinctness of cell borders in phase contrast images.
• Software-derived cell density measurements closely correlated with manual cell counting.
• Morphometric parameters (coefficient of variation, polygonality) were significantly correlated with cell density.

Conclusions:

• The developed software provides reliable morphometric data from HCEC phase contrast images.
• This software enables subjective and non-invasive quality assessment for corneal endothelium tissue engineering.
• The tool supports standardization and quality control in regenerative medicine for corneal diseases.