3D Biocomposite Culture Enhances Differentiation of Dopamine-like Neurons from SH-SY5Y cells: A Model for Studying Parkinson’s Disease Phenotypes

Studies of underlying neurodegenerative processes in Parkinson’s Disease (PD) have traditionally utilized cell cultures grown on two-dimensional (2D) surfaces. Biomimetic three-dimensional (3D) cell culture platforms have been developed to better emulate features of the brain’s natural microenvironment. We here use our bioengineered brain-like tissue model, composed of a silk-hydrogel composite, to study the 3D microenvironment’s contributions on the development and performance of dopaminergic-like neurons (DLNs). Compared with 2D culture, SH-SY5Y cells differentiated in 3D microenvironments were enriched for DLNs concomitant with a reduction in proliferative capacity during the neurodevelopmental process. Additionally, the 3D DLN cultures were more sensitive to oxidative stresses elicited by the PD-related neurotoxin 1-methyl-4-phenylpyridinium (MPP). MPP induced transcriptomic profile changes specific to 3D-differentiated DLN cultures, replicating the dysfunction of neuronal signaling pathways and mitochondrial dynamics implicated in PD. Overall, this physiologically-relevant 3D platform resembles a useful tool for studying dopamine neuron biology and interrogating molecular mechanisms underlying neurodegeneration in PD. Comparative gene expression profiling analysis of RNA-seq data for SH-SY5Y cells differentiated into dopamine-like neuron (DLN) cultures for 15 days using traditional 2D culture dishes and our bioengineered 3D silk-collagen platform. On day 13, 2D- and 3D-differentiated DLN cultures were treated with 1mM 1-methyl-4-phenylpyridinium (MPP) for 48 hours to investigate the effects of culture microenvironment on replicated features of PD.