Differentiation of human pluripotent stem cells (hPSCs) into neuronal cell types has become a cornerstone of neurological research by providing unique cellular models of human diseases. Lentiviral delivery of the transcription factor neurogenin-2 (NGN2) has been widely adopted over the past decade to rapidly convert hPSCs into neurons. However, this method faces challenges such as genomic integration risks, variability, labor-intensive steps and viral safety requirements.

To address these limitations we developed the STEMdiff™-TF Forebrain Induced Neuron Differentiation Kit, an integration-free system that simplifies the generation of highly-pure hPSC-derived neurons using mRNA-lipid nanoparticle (LNP) delivery of NGN2. Efficient neuron conversion can be achieved within 6 days, with 96% ± 3% of cells expressing neuronal marker TUJ1 (mean ± SD; n = 8, 4 cell lines). After two weeks of maturation using BrainPhys™ hPSC Neuron Kit, 94% ± 2.6% of neurons expressed MAP2, and 90% ± 4.9% expressed the synaptic vesicle marker synapsin I (n = 8, 4 cell lines). Transcriptomic analysis revealed a mostly glutamatergic neuron identity, with enrichment for genes SLC17A6 (vGLUT2) and GLS. In summary this kit provides an efficient, rapid and reproducible tool for generating functional glutamatergic neurons for neurobiological research.