Use of Levitated Cells n3D BiOAssay System to Determine Cytotoxicity of Calcium Sulfide (CaS) Clusters in Malignant Cells as Potential Cancer Cell Targeted Therapy (LB583)

Abstract

Preliminary results in our lab showed that naked Calcium Sulfide (CaS) nanoparticles (5.0nm) decreased cell proliferation in the carcinoma cell lines CRL‐2124. Objective: The objective of this study was to determine if CaS clusters measuring < 3.0 nm were cytotoxic to malignant cells when compared to benign fibroblasts CRL‐2522 by using the n3D BiOAssay (Biosciences, Inc.). Methods: A total of 100,000 cells per cell line were seeded in T‐75 flasks. When reaching 80% confluence, they were incubated with the n3D nano‐shuttles overnight (ON) at 37oC, 5% CO2, and 98% relative humidity. Next day, cells were trypsinized, counted, and seeded (1.4 ‐ 1.6 M/per well) in 6‐well plates using the n3D BiOAssay to levitate the cells. A total of 150,000 cells/well were seeded in a 96‐well plate in triplicates with a total volume of 300ul (150ul of cells, 120ul of media, and 30ul of the treatments). The treatments consist of 10 pmoles of CaS, 2% DMSO, or growth media. Analysis for cytotoxicity were performed with the ImageJ software by measuring the “O” ring outer and inner diameters. Results: The cell were successfully levitated by the n3D BiOAssay System. The expected “O” ring was formed before the treatments were added. We observed changes in the diameters for the CaS treated wells. Conclusions: The n3D BiOAssay provides an excellent and cost effective system for testing cellular toxicity of compounds with the potential to be used as therapeutic agents. We found that malignant cells treated with CaS showed more cytotoxicity than benign normal cells. CaS clusters may represent a novel Cancer Cell Targeted Therapy. Grant Funding Source: NIH‐NIGMSR25GM096955, NIH‐R25GM088023