Background: We study dopaminergic neurons with specific emphasis on Parkinson’s disease. Our work involves implementing procedures for examining in vitro mouse primary dopaminergic neurons as well as dopaminergic neurons derived from human stem cells. Here we want to establish a continuous cell line model for fast and cost-effective production of human dopaminergic-like cells to be used in our studies. Aims: Differentiate SH-SY5Y cells into dopaminergic-like cells using Retinoic Acid. Optimize the differentiation protocol for highest yield of TH positive cells. Methods: Retinoic Acid is used to induce the differentiation of SH-SY5Y cells, together with reduction of FBS concentration in the media. Later on, cells were split and transferred on the coated plates and the media was changed to Neurobasal media with appropriate supplements. Cells are fixed with 4% PFA, stained with anti-MAP2 and anti-TH antibodies and imaged with Zeiss LSM 980 confocal microscope. Results: Microscope images are analyzed visually and significant increase in TH positive cells is observed. Conclusions: We managed to differentiate SH-SY5Y cells to dopaminergic-like cells expressing TH. Moreover, differentiated cells looked more neuron-like and had longer processes. Ongoing work to be included in the poster: We will further validate the SH-SY5Y-derived dopaminergic neuron (SY5Y-DA) culture prior use in our studies. Efficiency of conversion to TH+/MAP2+ cells will be assessed by immunohistochemistry. Maturation of SY5Y-DA cells will be assessed by qPCR, using dopaminergic maturation markers. Function of cells will be assessed by electrophysiology and expression of the key neurotransmitter dopamine.