Using LUHMES Cells as a Model System to Study Dopaminergic Neuron Cell Biology and Parkinson’s Disease
Society for Neuroscience Meeting 2014
Washington, DC, United StatesNovember 15, 2014
Dopaminergic neurons play significant roles in motor, reward, and motivational behavior related circuits throughout the brain. To date, there are few continuous in vitro models available to laboratories in research, industry, and academia for studies related to basic dopaminergic cell biology or high throughput screening. Here, we propose the use of a human model system, LUHMES cells (ATCC CRL-2927), to study dopaminergic neuron cell biology and Parkinson’s disease. LUHMES cells are neuronal precursors derived from fetal ventral mesencephalon. Neuronal differentiation is governed by the termination of v-myc expression using low levels of tetracycline. During our characterization, we found that tetracycline induced differentiation resulted in consistent neurite outgrowth in LUHMES cells within 2 to 4 hours. One day post differentiation, cells displayed similar morphology, with several long processes protruding from the cell soma. Growth cones were often observed in early differentiated cultures. Immunocytochemistry in early differentiated cultures (Days in vitro, DIV 2-3) revealed low-level expression of tyrosine hydroxylase; however, these levels were increased significantly by 7 DIV with many neurons expressing tyrosine hydroxylase. We also investigated dopamine transporter expression. Differentiated LUHMES cultures were positive for neuronal markers such as bIII tubulin and devoid of expression of traditional glial markers including GFAP and IBA-1. Both undifferentiated and differentiated LUHMES cells were easily transfected using basic eGFP constructs, although greater efficiencies were observed with the use of viral constructs. In summary, LUHMES cells are a suitable and robust in vitro model system for studying dopaminergic neuron cell biology and mechanisms underlying Parkinson’s disease.
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