Synapses are essential for facilitating the transmission of information between neurons and for executing neurophysiological processes. Following the exocytosis of neurotransmitters, the synaptic vesicle may quickly undergo endocytosis to preserve the structural integrity of the synapse. When converting adipose-derived stromal cells (ADSCs) into neurons, the ADSCs have already demonstrated comparable morphology, structure, and electrophysiological characteristics to neurons. Nevertheless, there is currently no published study on the endocytotic function of neurons that are produced from ADSCs. This study aimed to examine synaptic endocytosis in neurons derived from ADSCs by qualitatively and quantitatively analyzing the presence of Ap-2, Clathrin, Endophilin, Dynamin, and Hsc70, which are the key proteins involved in clathrin-mediated endocytosis (CME), as well as by using FM1-43 and cadmium selenide quantum dots (CdSe QDs). Additionally, single-cell RNA sequencing (scRNA-seq) was used to look at the levels of both neuronal markers and markers related to CME at the same time. The results of this study provide evidence that synapses in neurons produced from ADSCs have a role in endocytosis, mainly through the CME route.