Conventional monolayer cell cultures continue to be an inexpensive and highly accessible model of human disease that can be easily harnessed to study the molecular and cellular mechanisms of photodynamic therapy (PDT). In this communication, a collection of informative assays for conventional cell cultures are provided to determine (1) the photosensitizer uptake kinetics and localization, (2) the efficacy of PDT using metabolism- or protein-based quantification methods, (3) the effects of PDT and combination treatments on the cell cycle, (4) the cell death pathways induced by PDT, and (5) the extent of mitochondrial membrane permeabilization of PDT and photochemotherapy combinations. For each type of assay, examples from the recent literature are provided in which novel photosensitizers, their nanocarriers, and various PDT-based combination therapies are investigated. Together, these assays are examples of approaches by which monolayer cell cultures can be used as a simple yet robust and versatile model to investigate PDT.
Keywords: Cytotoxicity; Dosimetry; Flow cytometry; Fluorescence microscopy; In vitro; Metabolism; Photochemotherapy; Phototherapy; Viability.
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