PICLS with human cells - first high-throughput screening method with human assay identifies novel longevity compounds including 2,5-anhydro-D-mannitol that increase chronological lifespan

Abstract Gerontology research on anti-aging interventions with drugs could be an answer to age-related diseases, aiming at closing the gap between lifespan and healthspan(definition). Here, we present two methods for assaying chronological lifespan in human cells: 1) a version of the classical outgrowth assay with quantitative assessment of surviving cells and 2) a version of the PICLS method (propidium iodide fluorescent-based measurement of cell death). Both methods are fast, simple to conduct, cost-effective, produce quantitative data for further analysis and can be used with diverse human cell lines. Whereas the first method is ideal for validation and testing the post-intervention reproductive potential of surviving cells, the second method has true high-throughput screening potential. The new technologies were validated with known anti-aging compounds (2,5-anhydro-D-mannitol and mTOR(definition)-inhibiting drug studied for extending healthspan and lifespan." style="text-decoration:underline dotted; text-underline-offset:2px; cursor:help;">rapamycin(definition)). Using the HTS method, we screened a library of 162 chemical entities and identified three novel anti-aging compounds that extend the cellular longevity of human cells.