Alzheimer’s disease (AD) is a progressive neurodegenerative disorder that causes problems with memory, thinking, and behavior. Currently, there are no medications available to prevent or eliminate the pathological phenomena of this disease, but symptomatic treatment is used, which can improve the patient’s condition. In this context, the aim of our study was to investigate the effect of roscovitine, a selective inhibitor of the neuron-specific kinase CDK5, on the viability of hippocampal neuronal cultures. CDK5 plays an important role in maintain- ing the structure and function of neurons, including synaptic plasticity and cytoskeletal integrity. Therefore, the study of the neuroprotective effect of roscovitine opens the way for its potential application in the treatment of neurodegenerative diseases, particularly AD. The study was conducted on rat hippocampal neuron cultures using double staining of cells with two DNA-binding dyes (Hoechst 33258 and propidium iodide). Neuronal viability was assessed via confocal laser scanning microscopy by comparing cell fluorescence under control conditions and after 24 hours of incubation with roscovitine. The results showed that roscovitine had minimal effects on neurons viability under normal conditions, causing only a slight increase in apoptosis and necrosis compared to controls, indicating its low toxicity on cells. Thus, our study highlights the importance of evaluating roscovitine’s effects for use in a cellular model of AD and determining its potential to counteract amyloid-beta-induced neurodegeneration