Darvishi Marzieh, Ghasemi Hamidabadi Hatef, Sahab Negah Sajad, Moayeri Ardeshir, Tiraihi Taki, Mirnajafi-Zadeh Javad, Jahanbazi Jahan-Abad Ali, Shojaei Amir
Department of Anatomy, Faculty of Medicine, Ilam University of Medical Sciences, Ilam, Iran.
Shefa Neuroscience Research Center, Khatam Alanbia Hospital, Tehran, Iran.
Iran J Basic Med Sci. 2020 Apr;23(4):431-438. doi: 10.22038/ijbms.2020.39797.9434.
Cell therapy has provided clinical applications to the treatment of motor neuron diseases. The current obstacle in stem cell therapy is to direct differentiation of stem cells into neurons in the neurodegenerative disorders. Biomaterial scaffolds can improve cell differentiation and are widely used in translational medicine and tissue engineering. The aim of this study was to compare the efficiency of two-dimensional with a three-dimensional culture system in their ability to generate functional motor neuron-like cells from adipose-derived stem cells.
We compared motor neuron-like cells derived from rat adipose tissue in differentiation, adhesion, proliferation, and functional properties on two-dimensional with three-dimensional culture systems. Neural differentiation was analyzed by immunocytochemistry for immature (Islet1) and mature (HB9, ChAT, and synaptophysin) motor neuron markers.
Our results indicated that the three-dimensional environment exhibited an increase in the number of Islet1. In contrast, two-dimensional culture system resulted in more homeobox gene (HB9), Choline Acetyltransferase (ChAT), and synaptophysin positive cells. The results of this investigation showed that proliferation and adhesion of motor neuron-like cells significantly increased in three-dimensional compared with two-dimensional environments.
The findings of this study suggested that three-dimension may create a proliferative niche for motor neuron-like cells. Overall, this study strengthens the idea that three-dimensional culture may mimic neural stem cell environment for neural tissue regeneration.
细胞疗法已在运动神经元疾病的治疗中得到临床应用。干细胞疗法当前的障碍在于如何在神经退行性疾病中将干细胞定向分化为神经元。生物材料支架可改善细胞分化,在转化医学和组织工程中广泛应用。本研究的目的是比较二维和三维培养系统从脂肪来源干细胞生成功能性运动神经元样细胞的效率。
我们比较了大鼠脂肪组织来源的运动神经元样细胞在二维和三维培养系统中的分化、黏附、增殖及功能特性。通过免疫细胞化学分析未成熟(胰岛1)和成熟(HB9、胆碱乙酰转移酶和突触素)运动神经元标志物来分析神经分化情况。
我们的结果表明,三维环境中胰岛1的数量有所增加。相比之下,二维培养系统产生了更多的同源框基因(HB9)、胆碱乙酰转移酶(ChAT)和突触素阳性细胞。本研究结果表明,与二维环境相比,三维环境中运动神经元样细胞的增殖和黏附显著增加。
本研究结果表明,三维环境可能为运动神经元样细胞创造一个增殖微环境。总体而言,本研究强化了三维培养可能模拟神经干细胞环境以促进神经组织再生的观点。