Merolli Antonio, Mao Yong, Voronin Gregory, Steele Joseph A M, Murthy N Sanjeeva, Kohn Joachim
New Jersey Center for Biomaterials, Rutgers - The State University of New Jersey, Piscataway, NJ, USA.
In Vivo Research Services, Rutgers - The State University of New Jersey, Piscataway, NJ, USA.
Neural Regen Res. 2019 Jun;14(6):1052-1059. doi: 10.4103/1673-5374.250626.
Information from the brain travels back and forth along peripheral nerves in the form of electrical impulses generated by neurons and these impulses have repetitive patterns. Schwann cells in peripheral nerves receive molecular signals from axons to coordinate the process of myelination. There is evidence, however, that non-molecular signals play an important role in myelination in the form of patterned electrical impulses generated by neuronal activity. The role of patterned electrical impulses has been investigated in the literature using co-cultures of neurons and myelinating cells. The co-culturing method, however, prevents the uncoupling of the direct effect of patterned electrical impulses on myelinating cells from the indirect effect mediated by neurons. To uncouple these effects and focus on the direct response of Schwann cells, we developed an in vitro model where an electroconductive carbon fiber acts as an artificial axon. The fiber provides only the biophysical characteristics of an axon but does not contribute any molecular signaling. In our "suspended wire model", the carbon fiber is suspended in a liquid media supported by a 3D printed scaffold. Patterned electrical impulses are generated by an Arduino 101 microcontroller. In this study, we describe the technology needed to set-up and eventually replicate this model. We also report on our initial in vitro tests where we were able to document the adherence and ensheath of human Schwann cells to the carbon fiber in the presence of patterned electrical impulses (hSCs were purchased from ScienCell Research Laboratories, Carlsbad, CA, USA; ScienCell fulfills the ethic requirements, including donor's consent). This technology will likely make feasible to investigate the response of Schwann cells to patterned electrical impulses in the future.
大脑发出的信息以神经元产生的电脉冲形式沿外周神经来回传输,这些脉冲具有重复模式。外周神经中的施万细胞接收来自轴突的分子信号以协调髓鞘形成过程。然而,有证据表明,非分子信号以神经元活动产生的模式化电脉冲形式在髓鞘形成中起重要作用。文献中使用神经元与髓鞘形成细胞的共培养研究了模式化电脉冲的作用。然而,共培养方法会阻止模式化电脉冲对髓鞘形成细胞的直接作用与神经元介导的间接作用解耦。为了解耦这些作用并聚焦于施万细胞的直接反应,我们开发了一种体外模型,其中导电碳纤维充当人工轴突。该纤维仅提供轴突的生物物理特性,不产生任何分子信号。在我们的“悬丝模型”中,碳纤维悬浮在由3D打印支架支撑的液体介质中。模式化电脉冲由Arduino 101微控制器产生。在本研究中,我们描述了建立并最终复制该模型所需的技术。我们还报告了我们最初的体外测试,在有模式化电脉冲的情况下,我们能够记录人类施万细胞对碳纤维的黏附与包裹(人类施万细胞购自美国加利福尼亚州卡尔斯巴德的ScienCell研究实验室;ScienCell符合伦理要求,包括捐赠者的同意)。这项技术未来可能会使研究施万细胞对模式化电脉冲的反应变得可行。