Developmental deficits and early signs of neurodegeneration revealed by PD patient derived dopamine neurons.

Abstract

Parkinson's disease (PD) is the second most common neurodegenerative disease affecting millions of elder people due to the degeneration of dopamine neurons in the striatum and substantia nigra. The clinical manifestations of PD include tremor, rigidity, bradykinesia and postural instability. Studying PD is challenging due to two obstacles: 1) disease models such as primary neurons or animal models usually couldn't recapitulate the disease phenotype, and 2) accessibility of human autopsied brain samples is very limited if not impossible. Induced pluripotent stem cells (iPSCs)-derived neuronal cells from patients emerge as an ideal in vitro model for disease modeling and drug development. Here we describe a cell density-dependent method for preparing functional hiPSC-derived dopamine neurons (iDAs) with ~90% purity (TH-positive cells). iDAs derived from PD patient exhibit the disease-related phenotypes, for example, slowed morphogenesis, reduced dopamine release, impaired mitochondrial function, and α-synuclein accumulation as early as 35days after induction. Furthermore, we found that the effects of cell density are different between iDA development stages, whereas high cell density increases stress for early neural progenitor cells (NPCs), but are neural-protective for mature iDAs, high density also favors morphogenesis. Hence, using stage and density-dependent strategies we can obtain high quality iDAs, which are critical for disease modeling, drug development and cell replacement therapy.