An MRC-5 cell based high-throughput, high-content imaging assay to identify hits against Trypanosoma cruzi intracellular parasites

by Zulfiqar B, Sykes ML, Escudié FB, Chatelain E, Avery VM. SLAS Discovery 2026,100298. doi: 10.1016/j.slasd.2026.100298

Summary: The discovery of effective therapies for Trypanosoma cruzi, the causative agent of Chagas disease, remains one of the most pressing challenges in parasitology and global health. Development of new therapies requires innovative and physiologically relevant assay platforms for high-throughput identification of compounds active against T. cruzi. The authors of this manuscript discuss the development, optimization, evaluation, and validation of a robust and highly reproducible high-throughput, high content imaging assay in a 384-well microplate format to quantitatively assess the effects of compounds on intracellular T. cruzi amastigotes infecting MRC-5 human lung fibroblasts. The multiplexed assay design enables concurrent evaluation of compound-induced cytotoxicity on host cells within the same well, serving as an early indicator of host cell viability and compound selectivity.

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