Adipose-derived mesenchymal stem cells are a great alternative to other types of stem cells obtained from other tissues, since they are found in large numbers and are easy to obtain by liposuction and do not have ethical connotations. These cells can differentiate into neuronal lineage using induction media, although the efficacy of these media is determined by their composition. In previous studies, we have demonstrated the differentiation of human adipose-derived mesenchymal stem cells to neuronal lineage by using three induction media, Neu1, Neu2 and Neu3, each one showing a series of neuronal markers in the treated cells. In the present study, a further step is taken, since once a neuronal differentiation is obtained with these three induction media, they are removed from the medium and both morphology and neuronal markers and the ability to maintain neurosphere formation are analyzed. The results obtained show that when these induction media are withdrawn in the three cases, both the neuronal morphological characteristics and the neuronal markers are lost at different times depending on the medium used. In the case of neurospheres, the ability to maintain this shape is also lost when they are not in contact with neuronal induction factors. Therefore, although the neuronal differentiation mechanism is very promising in this type of cells, it is necessary to carry out more studies to elucidate an induction medium that allows cells differentiated into neurons to maintain neuronal characteristics over time without the need to continue applying neuronal differentiation factors.