Cellular and Molecular Research Center, Basic Health Sciences Institute, Shahrekord University of Medical Sciences, Shahrekord, Iran.
Thalassemia and Hemoglobinopathy Research Center, Health Research Institute, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.
Int J Neurosci. 2020 Feb;130(2):107-116. doi: 10.1080/00207454.2019.1664518. Epub 2019 Oct 10.
With regard to their ease of harvest and common developmental origin, dental pulp stem cells (DPSCs) may act as a favorable source of stem cells in generation of nerves. Moreover; cellular migration and differentiation as well as survival, self-renewal, and proliferation of neuroprogenitor species require the presence of the central nervous system (CNS) mitogens including EGF and bFGF. Accordingly, the possibility of the induction of neuronal differentiation of DPSCs by EGF and bFGF was evaluated in the present study. DPSCs were treated with 20 ng/ml EGF, 20 ng/ml bFGF, and 10 µg/ml heparin. In order to further induce the neuroprogenitor differentiation, DPSC-derived spheres were also incubated in serum-free media for three days. The resulting spheres were then cultured in high-glucose Dulbecco's Modified Eagle Medium (DMEM) with 10% FBS. The morphology of the cells and the expression of the differentiation markers were correspondingly analyzed by quantitative polymerase chain reaction (qPCR), western blotting, and immunofluorescence (IF). The EGF/bFGF-treated DPSCs showed significant increase in the expression of the neuroprogenitor markers of Nestin and SRY (sex determining region Y)-box 2 (SOX2), 72 h after treatment. The up-regulation of Nestin and SOX2 induced by growth factors was confirmed using western blotting and IF. The cultures also yielded some neuron-like cells with a significant rise in Nestin, microtubule-associated protein 2 (MAP2), and Neurogenin 1 (Ngn1) transcript levels; compared with cells maintained in the control media ( < 0.05). DPSCs seemed to potentially differentiate into neuron-like cells under the herein-mentioned treatment conditions.
就其易于收获和共同的发育起源而言，牙髓干细胞（DPSCs）可能是生成神经的干细胞的有利来源。此外，细胞迁移和分化以及神经祖细胞的存活、自我更新和增殖都需要中枢神经系统（CNS）有丝分裂原的存在，包括表皮生长因子（EGF）和碱性成纤维细胞生长因子（bFGF）。因此，本研究评估了 EGF 和 bFGF 诱导 DPSCs 神经元分化的可能性。用 20ng/ml EGF、20ng/ml bFGF 和 10μg/ml 肝素处理 DPSCs。为了进一步诱导神经祖细胞分化，还将 DPSC 衍生的球体在无血清培养基中孵育三天。然后将得到的球体在高葡萄糖 DMEM 中培养，其中含有 10% FBS。通过定量聚合酶链反应（qPCR）、western blot 和免疫荧光（IF）相应地分析细胞形态和分化标志物的表达。EGF/bFGF 处理的 DPSCs 在处理后 72 小时，神经祖细胞标志物巢蛋白和性别决定区 Y 盒 2（SOX2）的表达显著增加。用 western blot 和 IF 证实了生长因子诱导的巢蛋白和 SOX2 的上调。与在对照培养基中维持的细胞相比（<0.05），培养物还产生了一些具有显著升高的巢蛋白、微管相关蛋白 2（MAP2）和神经生成素 1（Ngn1）转录水平的神经元样细胞。在所述处理条件下，DPSCs 似乎有可能分化为神经元样细胞。
