Treatment and measurement of metals in brain cell cultures

Metals play indispensable roles in a broad range of biological processes, ranging from catalyzing enzyme activities, cell signaling pathways, and maintaining protein structure and functions. While the distribution of metals is normally tightly regulated by various metalloproteins, accumulating evidence shows that metal dyshomeostasis plays significant roles in various health complications and diseases. As a therapeutic approach, studies have also explored the use of metal-bound compounds as potential therapeutic agents. In vitro screening is therefore crucial for the understanding of metal-regulating properties of readily available and novel metal-bound compounds. This protocol outlines steps involved in the treatment of mixed glia cultures with metal-bound compounds prior to further sample analysis, applied in the context of neurodegenerative disease. The protocol can also be optimized appropriately for application to other cell types and biological contexts. The protocol is compatible for downstream analyses including inductively coupled plasma mass spectrometry (ICP-MS), immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), gene expression and protein analyses, and X-ray fluorescence microscopy (XFM).