Research Center for Applied Sciences, Academia Sinica.
Research Center for Applied Sciences, Academia Sinica; Institute of Biophotonics, National Yang Ming Chao Tung University; Department of Mechanical and Mechatronic Engineering, National Taiwan Ocean University; College of Engineering, Chang Gung University;
J Vis Exp. 2021 Apr 14(170). doi: 10.3791/61917.
Physiological electric fields (EF) play vital roles in cell migration, differentiation, division, and death. This paper describes a microfluidic cell culture system that was used for a long-term cell differentiation study using microscopy. The microfluidic system consists of the following major components: an optically transparent electrotactic chip, a transparent indium-tin-oxide (ITO) heater, a culture media-filling pump, an electrical power supply, a high-frequency power amplifier, an EF multiplexer, a programmable X-Y-Z motorized stage, and an inverted phase-contrast microscope equipped with a digital camera. The microfluidic system is beneficial in simplifying the overall experimental setup and, in turn, the reagent and sample consumption. This work involves the differentiation of neural stem and progenitor cells (NPCs) induced by direct current (DC) pulse stimulation. In the stem cell maintenance medium, the mouse NPCs (mNPCs) differentiated into neurons, astrocytes, and oligodendrocytes after the DC pulse stimulation. The results suggest that simple DC pulse treatment could control the fate of mNPCs and could be used to develop therapeutic strategies for nervous system disorders. The system can be used for cell culture in multiple channels, for long-term EF stimulation, for cell morphological observation, and for automatic time-lapse image acquisition. This microfluidic system not only shortens the required experimental time, but also increases the accuracy of control on the microenvironment.
生理电场 (EF) 在细胞迁移、分化、分裂和死亡中发挥着重要作用。本文描述了一种微流控细胞培养系统,该系统用于使用显微镜进行长期细胞分化研究。微流控系统由以下主要组件组成:光学透明的趋电性芯片、透明的铟锡氧化物 (ITO) 加热器、培养基填充泵、电源、高频功率放大器、EF 多路复用器、可编程 X-Y-Z 电动台和配备数字摄像头的倒置相差显微镜。微流控系统有助于简化整体实验设置,并相应地减少试剂和样品的消耗。本工作涉及直流 (DC) 脉冲刺激诱导的神经干细胞和祖细胞 (NPCs) 的分化。在干细胞维持培养基中,经 DC 脉冲刺激后,小鼠 NPCs (mNPCs) 分化为神经元、星形胶质细胞和少突胶质细胞。结果表明,简单的 DC 脉冲处理可以控制 mNPCs 的命运,并可用于开发神经系统疾病的治疗策略。该系统可用于多通道细胞培养、长期 EF 刺激、细胞形态观察和自动定时图像采集。这种微流控系统不仅缩短了所需的实验时间,而且提高了对微环境的控制精度。