Optimizing resazurin-based viability assays for P-MSC/TER308 cell line to enhance results reliability

Objective: The results of this research contribute to the LifeSaver project, which focuses on reducing neonatal and infant mortality resulting from preterm births. The project aims to create an in vitro system simulating prenatal conditions to screen and analyze chemicals and pharmaceuticals, establishing scientifically justified regulations for their use during pregnancy. Because several papers have recently identified data inconsistencies in pre-clinical studies, a key part of the project involves optimizing cellular cytotoxicity assays to enhance the reliability of pharmacological and toxicity screening for drugs and environmental contaminants. Results: The resazurin-based viability assay was chosen as the primary method due to its widespread adoption and simplicity in assessing drug cytotoxicity. This work describes the optimization of the resazurin-based viability assay on the P-MSC/TERT308 cell line, a placenta-derived mesenchymal stem cell used within the LifeSaver project. By applying our previously described and validated Standard Operating Procedure, we fine-tuned experimental parameters, consistently obtaining reliable results with measurement uncertainty of less than 10%.