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A Non-destructive, Image Analysis Method for Evaluating Pigmentation in Induced Pluripotent Stem Cell-Derived Retinal
Gopika S Kumar1, Gowri P Chidambaranathan1, Anwar A Palakkan2
1Immunology & Stem Cell Biology, Aravind Medical Research Foundation, Madurai, Tamil Nadu, India.
Methods in Molecular Biology (Clifton, N.J.)
|April 30, 2025
Summary
We created an ImageJ macro to measure pigmentation in human induced pluripotent stem cell-derived retinal pigment epithelial cells. This method aids in screening compounds affecting pigmentation for potential therapeutic applications.
Area of Science:
- Cell Biology
- Biotechnology
- Ophthalmology
Background:
- Retinal pigment epithelial (RPE) cells are crucial for retinal health.
- Dyspigmentation in RPE cells is implicated in various eye diseases.
- Accurate assessment of RPE cell pigmentation is vital for research and drug discovery.
Purpose of the Study:
- To develop and validate an ImageJ macro for quantifying pigmentation in iPSC-derived RPE cells.
- To establish a reliable method for analyzing hyperpigmentation induced by hydroquinone.
- To provide a tool for screening compounds that modulate RPE cell pigmentation.
Main Methods:
- Cultured human induced pluripotent stem cell-derived RPE cells.
- Induced hyperpigmentation using hydroquinone and used untreated cells as controls.
- Analyzed bright-field images using a custom ImageJ macro.
- Validated image analysis results via spectrophotometric melanin quantification.
Main Results:
- Successfully developed an ImageJ macro for objective pigmentation analysis.
- Demonstrated the macro's ability to quantify hydroquinone-induced hyperpigmentation.
- Image analysis results correlated well with spectrophotometric melanin measurements.
Conclusions:
- The developed ImageJ macro offers a robust method for assessing pigmentation in iPSC-derived RPE cells.
- This tool can accelerate the screening of drugs and compounds affecting RPE cell pigmentation.
- Facilitates research into diseases associated with RPE dyspigmentation and potential treatments.
