The history of addiction therapeutics such as buprenorphine is rooted in attempts to develop non-addictive analgesics. Buprenorphine is used as an analgesic and as a medication assisted treatment to treat patients with opioid dependency. Buprenorphine is a semisynthetic opioid drug with partial agonistic properties that blocks and activates μ-opioid receptors. Studies suggest that the substance may have adverse effects on cell viability. Reduced neurogenesis and induction of apoptosis were observed in these studies. Using MTT, LDH and Calcein AM-assay, this experiment aims to evaluate the effects that buprenorphine has on cell viability of differentiated SH-SY5Y cells, which mimic neurons in the brain. First, the SH-SY5Y cells went through a growth phase, then a differentiation phase, followed by a treatment. SH-SY5Y cells were exposed to different concentrations of buprenorphine diluted in dimethyl sulfoxide (DMSO) and neural basal medium (NBM). Different cell markers were used in order to measure cell viability after the treatment. Visualization of the cells was achieved with Calcein AM and Hoechst stain. Statistical analyses to compile results were then performed with the statistic tool GraphPad Prism. Data collected from this study indicates that buprenorphine in high concentrations exhibit significant toxic effects on differentiated SH-SY5Y neural cells after only 24 hours of exposure.