en Cellular and molecular Neuroscience Cellular and molecular Neuroscience Original Original <p style="margin: 0px; text-align: justify; line-height: 200%; unicode-bidi: embed; direction: ltr;"><strong>Introduction</strong>: Cell therapy has been widely considered as a therapeutic approach for neurodegenerative diseases and nervous system damage. Cholinergic neurons as one of the most important neurons that play a significant role in controlling emotions, mobility, and autonomic systems. In this study, human dental pulp stem cells (hDPSCs) were differentiated into the cholinergic neurons by &beta;-mercaptoethanol in the preinduction phase and also by the nerve growth factor (NGF) in the induction phase.&nbsp;<br> <strong>Methods</strong>: The hDPSCs were evaluated for CD73, CD31, CD34, and Oct-4. Concentration-time relationships for NGF were assessed by evaluating the viability rate of cells and the immune response to nestin, neurofilament 160, microtubule-associated protein-2, and choline acetyltransferase.<br> <strong>Results</strong>: The hDPSCs had a negative response to CD34 and CD31. The optimal dose for the NGF was 50 ng/mL seven days after the induction when the highest percentage of expressing markers for the cholinergic neurons (ChAT) was detected.<br> <strong>Conclusion</strong>: The results of this study provided a method for producing cholinergic neurons by hDPSCs, which can be used in cytotherapy for degenerative diseases of the nervous system and also spinal cord injury.</p> Dental pulp, Stem cells, Cholinergic neurons, Differentiation, Nerve growth factor 609 618 http://bcn.iums.ac.ir/browse.php?a_code=A-10-973-2&slc_lang=en&sid=1 Shahram Darabi 13700319475328460025367 13700319475328460025367 No Cellular and Molecular Research Center, Qazvin University of Medical Science, Qazvin, Iran. Taki Tiraihi 13700319475328460025368 13700319475328460025368 No Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran. Maryam Nazm Bojnordi 13700319475328460025369 13700319475328460025369 No Department of Anatomy & Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran. Hatef Ghasemi Hamidabadi hatefdr@gmail.com 13700319475328460025370 13700319475328460025370 Yes Department of Anatomy & Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran.; Immunogenetic Research Center, Department of Anatomy & Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran. Nourollah Rezaei 13700319475328460025371 13700319475328460025371 No Department of Anatomy & Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran.; Immunogenetic Research Center, Department of Anatomy & Cell Biology, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran. Maria Zahiri 13700319475328460025372 13700319475328460025372 No The Persian Gulf Marine Biotechnology Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran.; Department of Anatomical Sciences, School of Medical Sciences, Bushehr University of Medical Sciences, Bushehr, Iran. Rafieh Alizadeh 13700319475328460025373 13700319475328460025373 No ENT and Head & Neck Research Center and Department, Hazrat Rasoul Akram Hospital, Iran University of Medical Sciences, Tehran, Iran.