Darabi Shahram, Tiraihi Taki, Nazm Bojnordi Maryam, Ghasemi Hamidabadi Hatef, Rezaei Nourollah, Zahiri Maria, Alizadeh Rafieh
Cellular and Molecular Research Center, Qazvin University of Medical Science, Qazvin, Iran.
Department of Anatomical Sciences, Faculty of Medical Sciences, Tarbiat Modares University, Tehran.
Basic Clin Neurosci. 2019 Nov-Dec;10(6):609-617. doi: 10.32598/bcn.10.6.609. Epub 2019 Nov 1.
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 β-mercaptoethanol in the preinduction phase and also by the nerve growth factor (NGF) in the induction phase.
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.
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.
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.
细胞疗法已被广泛视为治疗神经退行性疾病和神经系统损伤的一种治疗方法。胆碱能神经元是在控制情绪、运动和自主神经系统中发挥重要作用的最重要神经元之一。在本研究中，人牙髓干细胞（hDPSCs）在预诱导阶段通过β-巯基乙醇以及在诱导阶段通过神经生长因子（NGF）分化为胆碱能神经元。
对hDPSCs进行CD73、CD31、CD34和Oct-4评估。通过评估细胞活力率以及对巢蛋白、神经丝160、微管相关蛋白-2和胆碱乙酰转移酶的免疫反应来评估NGF的浓度-时间关系。
hDPSCs对CD34和CD31呈阴性反应。诱导7天后，NGF的最佳剂量为50 ng/mL，此时检测到胆碱能神经元（ChAT）表达标志物的百分比最高。
本研究结果提供了一种通过hDPSCs产生胆碱能神经元的方法，可用于神经系统退行性疾病以及脊髓损伤的细胞治疗。
