Abstract

Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease that primarily affects upper and lower motor neurons. The pathology of ALS associates closely with cellular level physiological abnormalities in TAR DNA binding protein (TDP-43), a ubiquitously expressed, highly conserved nuclear protein. TDP-43 proteinopathy involves excessive mislocalization and misfolding within the cytoplasm where certain components of the protein aggregate into hyper-phosphorylated tau-negative ubiquitinated inclusions. These intracellular alterations impair neurite growth during neuronal differentiation while damaging cell viability and ultimately causing apoptotic necrosis in affected tissues. Studying the effects of ALS induced TDP-43 aggregation in vitro requires the establishment of a neuronal cell culture that recapitulates the physiology of human motor neurons. NSC-34 cells can be differentiated to model selected aspects of motor neuron development. In this study, an NSC-34 cell culture system is established for the future comparison of cells transiently transfected for regulated expression of both wild-type and C-terminal mutated (Q311K) TDP-43 proteins. The results indicate that NSC-34 cells seeded onto a 96-well plate in serum-free media optimally differentiate motor neuron-like morphology, as measured by intra-well neurite length, when plated at 10k cells/well.

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