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Measuring Single-Cell Aging with an Imaging-based Biomarker of Chromatin and Epigenetic Aging

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Quantitative Cellular AGing Evaluation system (qCAGEs): a dual-parameter platform for high-throughput senescence

Dong-Ju Shin^1,2,3, Young-Sam Lee⁴, Chong Won Choi⁵

¹Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul, South Korea.

|June 26, 2026

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A new screening platform, quantitative Cellular AGing Evaluation system (qCAGEs), accurately identifies senescence-modulating compounds by measuring aging markers and cell viability simultaneously. This system aids in developing senolytic and anti-aging therapies.

Area of Science:

Cellular biology
Biotechnology
Pharmacology

Background:

Accurate identification of senescence-modulating compounds requires advanced screening.
Current methods using single parameters cannot differentiate specific drug responses from cytotoxicity.
Distinguishing senolytic and anti-aging effects necessitates multi-parameter analysis.

Purpose of the Study:

To develop a dual-parameter high-throughput screening platform for precise identification and quantification of senescence-modulating compounds.
To establish a standardized method for classifying drug responses based on cellular aging status.
To enable systematic phenotypic classification of drug responses in therapeutic development.

Main Methods:

Developed the quantitative Cellular AGing Evaluation system (qCAGEs) measuring α-L-fucosidase activity (4ME) and cell viability.

Keywords:

Anti-aging Cellular senescence High-throughput screening Three-dimensional culture α-L-fucosidase

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Established the Cell count-Aging activity Reference Line (CARL) for baseline relationship definition.

Validated the platform using human dermal fibroblast senescence models and 3D culture formats.

Main Results:

The 4ME/cell count ratio effectively discriminated senescent from young cells with a >sixfold difference.
Assay precision met high-throughput screening criteria (CV < 15%).
Platform adaptation to 3D cultures showed strong concordance with 2D results (cosine similarity > 0.5 for >75% of compounds).

Conclusions:

qCAGEs provides a standardized, quantitative approach for evaluating drug-induced changes in cellular aging.
The platform enables accurate classification of drug responses into anti-aging, pro-aging, senolytic, and cytotoxic categories.
qCAGEs has broad applications in senolytic and anti-aging therapeutic development.