Microtopography-Enhancement on Dopaminergic Neuronal Differentiation

Micro and nanotopograhy have been shown to be able direct stem cell fate. Substrate topography also has the potential in differentiating stem cells into subtype specific cells. Parkinson’s disease is a neurodegenerative disease attributed to the loss of midbrain dopaminergic (DA) neurons. Differentiation of stem cells into subtype specific cells may be guided by appropriate topographical cues but the role of topography has hitherto not been well understood. We aim to develop an in vitro substrate that will accelerate the derivation of midbrain dopaminergic neurons from human pluripotent stem cells (hPSCs) and neural progenitor cells.

Neuronal differentiation of murine hippocampal neural progenitor cells (mNPCs) on a Multi-Architecture (MARC) chip with various topographical structures was performed to identify the topographies that generated the highest percentage of neuronal (b-III-tubulin positive) and dopaminergic (tyrosine hydroxylase positive) populations. Subsequently, the ability of topographical patterns to influence the differentiation of hPSCs into subtype-specific and regionalized dopaminergic neurons was investigated. We have made minor modifications to the protocol based on dual SMAD inhibition method and optimized on patterned substrates to further improve the efficiency for dopaminergic neuron derivation.

From both type of stem cells, the topographical patterns could enhance the dopaminergic neuron differentiation. These results show the use of topographical influence for neuronal subtype specification, which could be translated into new therapeutic approaches for this neurodegenerative disease.