Gervois P, Wolfs E, Dillen Y, Hilkens P, Ratajczak J, Driesen R B, Vangansewinkel T, Bronckaers A, Brône B, Struys T, Lambrichts I
1 Morphology Research Group, Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium.
2 Group of Physiology, Biomedical Research Institute, Hasselt University, Diepenbeek, Belgium.
J Dent Res. 2017 Jun;96(6):654-662. doi: 10.1177/0022034517690491. Epub 2017 Jan 31.
Neurological disorders are characterized by neurodegeneration and/or loss of neuronal function, which cannot be adequately repaired by the host. Therefore, there is need for novel treatment options such as cell-based therapies that aim to salvage or reconstitute the lost tissue or that stimulate host repair. The present study aimed to evaluate the paracrine effects of human dental pulp stem cells (hDPSCs) on the migration and neural maturation of human SH-SY5Y neuroblastoma cells. The hDPSC secretome had a significant chemoattractive effect on SH-SY5Y cells as shown by a transwell assay. To evaluate neural maturation, SH-SY5Y cells were first induced toward neuronal cells, after which they were exposed to the hDPSC secretome. In addition, SH-SY5Y cells subjected to the hDPSC secretome showed increased neuritogenesis compared with nonexposed cells. Maturated cells were shown to increase immune reactivity for neuronal markers compared with controls. Ultrastructurally, retinoic acid (RA) signaling and subsequent exposure to the hDPSC secretome induced a gradual rise in metabolic activity and neuronal features such as multivesicular bodies and cytoskeletal elements associated with cellular communication. In addition, electrophysiological recordings of differentiating cells demonstrated a transition toward a neuronal electrophysiological profile based on the maximum tetrodotoxin (TTX)-sensitive, Na current. Moreover, conditioned medium (CM)-hDPSC-maturated SH-SY5Y cells developed distinct features including, Cd-sensitive currents, which suggests that CM-hDPSC-maturated SH-SY5Y acquired voltage-gated Ca channels. The results reported in this study demonstrate the potential of hDPSCs to support differentiation and recruitment of cells with neuronal precursor characteristics in a paracrine manner. Moreover, this in vitro experimental design showed that the widely used SH-SY5Y cell line can improve and simplify the preclinical in vitro research on the molecular mechanisms of stem cell-mediated neuronal regeneration.
神经疾病的特征是神经退行性变和/或神经元功能丧失，而宿主无法对其进行充分修复。因此，需要新的治疗方法，如旨在挽救或重建受损组织或刺激宿主修复的细胞疗法。本研究旨在评估人牙髓干细胞（hDPSC）对人SH-SY5Y神经母细胞瘤细胞迁移和神经成熟的旁分泌作用。如Transwell实验所示，hDPSC分泌组对SH-SY5Y细胞具有显著的趋化作用。为了评估神经成熟情况，先将SH-SY5Y细胞诱导为神经元细胞，然后使其暴露于hDPSC分泌组。此外，与未暴露的细胞相比，暴露于hDPSC分泌组的SH-SY5Y细胞神经突生成增加。与对照组相比，成熟细胞对神经元标志物的免疫反应性增强。在超微结构上，视黄酸（RA）信号传导以及随后暴露于hDPSC分泌组会导致代谢活性逐渐升高，并出现与细胞通讯相关的多泡体和细胞骨架成分等神经元特征。此外，对分化细胞的电生理记录表明，基于最大河豚毒素（TTX）敏感性钠电流，细胞向神经元电生理特征转变。此外，条件培养基（CM）-hDPSC-成熟的SH-SY5Y细胞表现出独特的特征，包括对镉敏感的电流，这表明CM-hDPSC-成熟的SH-SY5Y细胞获得了电压门控钙通道。本研究报告的结果表明，hDPSC具有以旁分泌方式支持具有神经元前体特征的细胞分化和募集的潜力。此外，这种体外实验设计表明，广泛使用的SH-SY5Y细胞系可以改善和简化关于干细胞介导的神经元再生分子机制的临床前体外研究。
