Determining Neuronal Cell Line Protocol for C60 Treatment

Abstract

Parkinson’s disease (PD) is characterized by the buildup of Reactive Oxygen Species (ROS) that cause oxidative stress on dopaminergic cells, ending in apoptosis of these neurons. Alzheimer ’s disease (AD) has two markers of its pathology: Neurofibrillary Tangles (NFTs) and Amyloid Plaque buildup. Amyloid Plaque causes the deterioration of neural pathways by clogging synapses and causing oxidative stress. Carboxyfullerene (C60) is a lipid-soluble, symmetric molecule that has been researched on its ability to take up radical electrons that are present in ROS. In this experiment, we aimed to establish a foundation for further exploration of water-soluble C60 in treating symptoms of PD and AD. For the purposes of making C60 water-soluble, a fullerene derivative of C60(C3H4O4)3 was utilized. PC12 cells were differentiated using Nerve Growth Factor (NGF) and were placed into 12 trials of test groups and controls. Test groups contained C60, H2O2, and PC12 cells with NGF. An AlamarBlue absorbance assay was the marker used for determining cell viability, along with visual inspection of cultures under an inverted microscope. The percent reduction values collected confounded by varying cell density in each well. This experiment successfully established a protocol for culturing neuronal cell lines with biocompatible nanoparticle treatment. Repetition of this experiment with the new protocols created, while adjusting for confounding variables, will lead to more conclusive results for understanding the applications of C60 on neuronal model