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Publication Detail

Title: Enhanced differentiation of human dopaminergic neuronal cell model for preclinical translational research in Parkinson's disease.

Authors: Harischandra, Dilshan S; Rokad, Dharmin; Ghaisas, Shivani; Verma, Saurabh; Robertson, Alan; Jin, Huajun; Anantharam, Vellareddy; Kanthasamy, Arthi; Kanthasamy, Anumantha G

Published In Biochim Biophys Acta Mol Basis Dis, (2020 04 01)

Abstract: Human-derived neuronal cell lines are progressively being utilized in understanding neurobiology and preclinical translational research as they are biologically more relevant than rodent-derived cells lines. The Lund human mesencephalic (LUHMES) cell line comprises human neuronal cells that can be differentiated to post-mitotic neurons and is increasingly being used as an in vitro model for various neurodegenerative diseases. A previously published 2-step differentiation procedure leads to the generation of post-mitotic neurons within 5-days, but only a small proportion (10%) of the total cell population tests positive for tyrosine hydroxylase (TH). Here we report on a novel differentiation protocol that we optimized by using a cocktail of neurotrophic factors, pleiotropic cytokines, and antioxidants to effectively generate proportionately more dopaminergic neurons within the same time period. Visualization and quantification of TH-positive cells revealed that under our new protocol, 25% of the total cell population expressed markers of dopaminergic neurons with the TH-positive neuron count peaking on day 5. These neurons showed spontaneous electrical activity and responded to known Parkinsonian toxins as expected by showing decreased cell viability and dopamine uptake and a concomitant increase in apoptotic cell death. Together, our results outline an improved method for generating a higher proportion of dopaminergic neurons, thus making these cells an ideal neuronal culture model of Parkinson's disease (PD) for translational research.

PubMed ID: 31442530 Exiting the NIEHS site

MeSH Terms: Antigens, Differentiation/biosynthesis; Cell Differentiation*; Cell Line; Dopaminergic Neurons/metabolism*; Dopaminergic Neurons/pathology; Humans; Models, Neurological*; Neural Stem Cells/metabolism*; Neural Stem Cells/pathology; Parkinson Disease/metabolism*; Parkinson Disease/pathology; Parkinson Disease/therapy; Translational Medical Research*

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