Aims Neurological disorders continue to be challenging to treat due to the limited capacity for neuronal regeneration in the central nervous system. Mesenchymal stem cells present a promising approach for neuron replacement therapies. This study aimed to establish a novel protocol for differentiating mesenchymal stem cells into Purkinje-like cells, given the critical role of Purkinje neurons in motor coordination, cognition, and emotional regulation. Materials & Methods Mesenchymal stem cells were isolated from the bone marrow of mice and characterized using specific markers (CD34, CD44, CD45, and CD73). Subsequently, the mesenchymal stem cells were differentiated into neural stem cells, which were characterized through immunocytochemistry and neurosphere ormation (Nestin and SOX2). Finally, neural stem cells were dierentiated into Purkinje-like cells using specific culture media and identified by morphology and immunocytochemistry (calbindin D28K and IP3R1). Findings According to the criteria set by the International Society for Cell and Therapy, isolated mesenchymal stem cells adhered to the flask surace and expressed the CD73 and CD44 markers but did not express the CD34 and CD45 markers. The identity o neural stem cells was confirmed through their morphology, expression o Nestin and SOX2 markers and neurosphere ormation. The markers calbindin D28K and IP3R1 were significantly expressed in Purkinje-like cells differentiated from mesenchymal stem cells. Conclusion The Purkinje-like cells created in this study can serve as an in vitro model to develop appropriate treatments for diseases caused by the dysfunction of Purkinje cells.

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