Cultured networks of excitatory projection neurons and inhibitory interneurons for studying human cortical neurotoxicity

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Science Translational Medicine  06 Apr 2016:
Vol. 8, Issue 333, pp. 333ra48
DOI: 10.1126/scitranslmed.aad0623

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Insights into neuronal cell death

The study of the mechanisms of cell death in human cortical neurons has been hampered by the lack of human neuronal cultures that exhibit a balanced network of excitatory and inhibitory synapses. Xu et al. now describe a method to culture human neurons with a representative ratio of both excitatory and inhibitory neurons derived from human embryonic stem cells or human inducible pluripotent stem cells. Using this new method, they show that human cortical neurons die in a nitric oxide– and poly(ADP-ribose) polymerase-1–dependent manner. These cultures can be used to study the mechanisms of neurotoxicity in human disorders that involve the demise of cortical neurons.


Translating neuroprotective treatments from discovery in cell and animal models to the clinic has proven challenging. To reduce the gap between basic studies of neurotoxicity and neuroprotection and clinically relevant therapies, we developed a human cortical neuron culture system from human embryonic stem cells or human inducible pluripotent stem cells that generated both excitatory and inhibitory neuronal networks resembling the composition of the human cortex. This methodology used timed administration of retinoic acid to FOXG1+ neural precursor cells leading to differentiation of neuronal populations representative of the six cortical layers with both excitatory and inhibitory neuronal networks that were functional and homeostatically stable. In human cortical neuronal cultures, excitotoxicity or ischemia due to oxygen and glucose deprivation led to cell death that was dependent on N-methyl-d-aspartate (NMDA) receptors, nitric oxide (NO), and poly(ADP-ribose) polymerase (PARP) (a cell death pathway called parthanatos that is distinct from apoptosis, necroptosis, and other forms of cell death). Neuronal cell death was attenuated by PARP inhibitors that are currently in clinical trials for cancer treatment. This culture system provides a new platform for the study of human cortical neurotoxicity and suggests that PARP inhibitors may be useful for ameliorating excitotoxic and ischemic cell death in human neurons.

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