Quick-Neuron™ Excitatory – Human iPSC-Derived Neurons (Healthy Donor)

Our easy-to-follow, optimized excitatory neuron protocol supports rapid and reproducible differentiation of human induced pluripotent stem cells (iPSCs). This step-by-step guide outlines key culture stages and timelines for generating functional excitatory neurons from human iPSCs.

Excitatory Neuron morphology is confirmed via phase contrast imagery. Representative phase contrast images of Quick-Neuron™ Excitatory – Human iPSC-derived Neurons on days 1-7 post-thaw (scale bars = 100 μm).

Rigorous post-differentiation characterization of our iPSC-derived excitatory neurons ensures identity, purity, and functional relevance. Using excitatory neuron–specific markers, phenotypic profiling, and functional assays, we confirm lineage specification, neuronal maturity, and batch-to-batch consistency.

Expression of excitatory neuron–specific markers is evaluated post-differentiation to verify glutamatergic identity, purity, and maturation status. Using transcript and protein-level analyses, we confirm robust expression of established cortical and excitatory lineage markers to ensure batch-to-batch consistency and experimental reliability.

Whole-Cell Patch Clamp

Electrophysiological properties of iPSC-derived excitatory neurons. Whole-cell patch clamp of Quick-Neuron™ Excitatory – Human iPSC-derived Neurons 5-6 weeks post-thaw. (A) A brightfield image of the neurons measured. (B) Spontaneous action potentials were recorded. (C) Spontaneous excitatory postsynaptic current of neurons was detected by voltage clamp measurement at -70mV, indicating the formation of mature synapses. Data courtesy of E-PHY SCIENCE SAS.

Multi-Electrode Array (MEA)

Excitatory Neurons seeded onto MEA plates remain functional after transport. An MEA plate seeded with Quick-Neuron™ Excitatory Neurons and primary human astrocytes was transported from Tokyo, Japan to Maryland, USA. The baseline network burst firing of one well was measured on day 49 post-thaw before transport (A) and on day 50 after 33-hr transport (B). (C) Network burst firing on day 51 exhibited responses to varying concentrations of 4-AP after transport. Data courtesy of Ricoh.

Transcriptomic Validation of Human iPSC-Derived Excitatory Neurons

RNA-Sequencing

Gene Expression of Quick-Neuron™ Excitatory Neurons. A heat map of selected gene expression data from RNA-seq performed on undifferentiated iPSCs, Quick-Neuron™ Excitatory Neurons (EX only) cultured for 10 days and 38 days, Quick-Neuron™ Excitatory Neurons cocultured with primary astrocytes (EX+AST) for 18 and 52 days, and primary fetal and adult brain samples is shown. Values represent log10(TPM + 1). Data courtesy of Ricoh.

Principal Component Analysis (PCA)

Quick-Neuron™ Excitatory Neurons display gene expression profiles similar to those of human brain: RNA-seq was performed on undifferentiated iPSCs, Quick-Neuron™Excitatory Neurons (EX only) cultured for 10 days and 38 days, Excitatory neurons cocultured with primary astrocytes (EX+AST) for 18 and 52 days, and primary fetal and adult brain samples. Principal component analysis indicates that Quick-Neuron™ Excitatory Neurons display gene expression similar to that of the human brain, particularly when grown with astrocytes. As cells remain in culture over time they more closely resemble adult human brain cells. Data courtesy of Ricoh.