The differentiation of functional cortical neurons from human induced pluripotent stem cells in vitro easily lends itself to a serum-free, drug delivery platform advantageous for testing novel chemicals for safety and efficacy in disease treatment. Initially, cortical neuron cultures were characterized morphologically by phase microscopy and immunocytochemistry and functionally by patch-clamp electrophysiology. Specifically, the expression of neuronal markers and neuronal activity increased throughout maturation. On day 0 of maturation, 50 percent of the culture expressed layer V cortical neuron marker ctip2 and neuronal marker beta-III tubulin and displayed spontaneous and repetitive firing through whole-cell patch clamp. By day 28 of maturation, 90 percent of the culture expressed the aforementioned markers and displayed electrical activity. Subsequently, neurons were cultured on multi-electrode arrays (MEAs) to determine the effects of chemicals on neural circuit physiology for modeling brain disease phenotypes. In this system, we tested GABAA receptor antagonists and agonists as chemical convulsants or anti-convulsants, respectively. GABAA receptor antagonist administration enhanced spontaneous activity mimicking an epileptic phenotype, while GABAA receptor agonist administration quieted spontaneous activity. The versatility of this model lies in its ability to present an array of brain diseases characterized by functional brain deficits. Chemicals affecting receptor binding can be added to either enhance or inhibit neuronal activity. This serum-free, hiPSC cortical neuron model establishes a platform for the evaluation of neuron activity as well as a platform for drug testing in vitro.

Development and characterization of hiPSC cortical neurons and their application to drug evaluation in CNS disease models

2019 Spring National Meeting

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