The Ion AmpliSeqTM was employed for the temporal transcriptomic characterization of the maturation of human stem cell derived neurons at multiple time points. Analysis of the data revealed a progressive increase in markers associated with neuronal development and astrocyte markers over the time. Indicating the establishment of a co-culture containing both glial cells and neurons, and highlighting the importance of culture duration. With the help of unbiased bioinformatics, the top 100 genes driving the principal component analysis clustering and the top 20 differentially expressed genes associated with neuronal maturation were identified. Alongside pathway enrichment analysis, these findings revealed that the neurons exhibited a more pronounced GABAergic phenotype, signifying their specialization towards this specific neuronal phenotype. These maturing neurons demonstrate a robust culture and progress through various stages of development. Later, these cells were used to identify neurodevelopmental effect of the specific serotonin reuptake inhibitors fluoxetine and paroxetine at therapeutically or clinically relevant concentrations through downstream analysis such as neurite outgrowth measurements. At a concentration of 10 µM fluoxetine, which is higher than plasma levels but lower than brain concentration, significantly led to a decline in cellular processes, cell body integrity and neurite branching. Conversely, paroxetine, administered at therapeutic plasma concentrations (0.05 and 0.2 µM), did not yield statistically significant results, although it demonstrated a noticeable trend of reduction in neurite outgrowth parameters. Ampliseq, RT-qPCR and uptake experiments revealed no detectable serotonin transporter (SERT) activity in this cell model. This emphasizes that the observed side effects are independent of SERT activity.