Here, we investigated the expression of genes associated with the differentiation of NPCs during three weeks in dopaminergic differentiation media. Known early neuron markers, MAP2 and Tuj1 genes reached peak expression at two weeks while expression of dopaminergic neuronal genes (TH, Nurr1, VMAT2, AADC) was significantly increased in a time-dependent manner (p < 0.05) in two types of normal NPCs (Figure 1). Furthermore, expression of genes associated with glutamatergic (vGLUT1, vGLUT2, GLS2; Figure 2) and GABAergic (GABRB3; Figure 3) neurons was also induced and peaked at the end of three weeks. This shows that ATCC NPCs and dopaminergic differentiation media are capable of producing GABAergic and glutamatergic neurons, in addition to dopaminergic neurons. Noticeably, there was significant induction of motor neuron gene (EN1, LIM3, Hb9) expression in ACS-5003™ NPCs while there was significant induction of cholinergic (ChAT) neuron gene expression in ACS-5007™ NPCs, but not in ACS-5003™ NPCs during dopaminergic differentiation (Figure 3). Moreover, expression of TH, Tuj1, Nurr1, GLS2, and vGLUT2 neuronal markers in both ACS-5003™- and ACS-5007™-derived neurons has been confirmed by immunocytochemistry (Figure 4). To validate that ATCC NPCs and dopaminergic neuron differentiation media are suitable for drug screening, we conducted neurotoxicity screenings in the two types of NPCs and NPC-derived neurons by using a resazurin viability assay and high-content imaging analysis. We found that paclitaxel, a microtubule-stabilizing chemotherapeutic agent, significantly induced neurotoxicity (p < 0.001) in the two types of NPCs evaluated at a IC50 of ~ 1 µM, but not in NPC-derived neurons (Figures 5 and 6). Vincristine, amiodarone, and chlorhexidine significantly decreased viability of both NPCs and neurons, whereas piperine, cisplatin, and hydroxyurea did not induce any significant neurotoxicity in neurons (Figures 5 and 7). This study demonstrates that ATCC iPSC-derived NPCs and dopaminergic differentiation media are suitable for studying neurological development and neurotoxicity screening.