An In Vitro Model of Human Dopaminergic Neurons Derived from Embryonic Stem Cells: MPP+ Toxicity and GDNF Neuroprotection

Abstract

Human embryonic stem cells (hESCs) can proliferate indefinitely yet also differentiate in vitro, allowing normal human neurons to be generated in unlimited numbers. Here, we describe the development of an in vitro neurotoxicity assay using human dopaminergic neurons derived from hESCs. We showed that the dopaminergic neurotoxin 1-methyl-4-phenylpyridinium (MPP + ), which produces features of Parkinson's disease in humans, was toxic for hESC-derived dopaminergic neurons. Treatment with glial cell line-derived neurotrophic factor protected tyrosine hydroxylase-positive neurons against MPP + -induced apoptotic cell death and loss of neuronal processes as well as against the formation of intracellular reactive oxygen species. The availability of human dopaminergic neurons, derived from hESCs, therefore allows for the possibility of directly examining the unique features of human dopaminergic neurons with respect to their responses to pharmacological agents as well as environmental and chemical toxins.

Dopaminergic Neurons Derived from Human Embryonic Stem Cell Derived Neural Progenitors: Biological Relevance and Application

“…The use of GDNF along with co-culture with PA6 cells increased the number of TH positive cells produced over PA6 co-culture alone (BuytaertHoefen et al, 2004). Using hESC derived dopaminergic neurons as a model of PD, GDNF provided protection against MPTP toxicity (Zeng et al, 2006). Another method of improving delivery systems involves genetically altering neural stem cells or astrocytes to release GDNF and injecting these into the brain which protected from parkinsonian motor responses in mouse models of PD (Engele andFranke, 1996, Elsworth et al, 2008).…”

Acrylamide alters neurotransmitter induced calcium responses in murine ESC-derived and primary neurons

“…Fritsche et al, 2005;Gassmann et al, 2010;Huang and Schneider, 2004;Tamm et al, 2006;Tateno et al, 2004;Zimmer et al, 2011Zimmer et al, , 2012. Up to now, these cell systems have hardly been used to analyze toxic effects on fully differentiated neurons, yet (Visan et al, 2012;Zeng et al, 2006); especially not, when toxins affect neuronal function. Such effects have been demonstrated using electrophysiological methods in cultures of primary neurons from rodents (Defranchi et al, 2011;Stett et al, 2003;van Vliet et al, 2007).…”

High-Throughput Transfection of Differentiated Primary Neurons from Rat Forebrain

“…The utility of in vitro models is directly related to accurate representation of the in vivo system they are designed to mimic. 1 Because of availability and convenience, immortalized cell lines are often employed instead of primary cells. In the case of neuronal studies, few cell lines can serve as accurate in vitro models of neurons.…”