Su Ping-Jung, Liu Zongbin, Zhang Kai, Han Xin, Saito Yuki, Xia Xiaojun, Yokoi Kenji, Shen Haifa, Qin Lidong
Department of Nanomedicine, Houston Methodist Research Institute, Houston, TX 77030, USA.
Department of Cell and Developmental Biology, Weill Medical College of Cornell University, New York, NY 10065, USA.
Sci Rep. 2015 Aug 28;5:13591. doi: 10.1038/srep13591.
In vitro culture of dissociated retinal neurons is an important model for investigating retinal synaptic regeneration (RSR) and exploring potentials in artificial retina. Here, retinal precursor cells were cultured in a microfluidic chip with multiple arrays of microchannels in order to reconstruct the retinal neuronal synapse. The cultured retinal cells were physically connected through microchannels. Activation of electric signal transduction by the cells through the microchannels was demonstrated by administration of glycinergic factors. In addition, an image-based analytical method was used to quantify the synaptic connections and to assess the kinetics of synaptic regeneration. The rate of RSR decreased significantly below 100 μM of inhibitor glycine and then approached to a relatively constant level at higher concentrations. Furthermore, RSR was enhanced by chemical stimulation with potassium chloride. Collectively, the microfluidic synaptic regeneration chip provides a novel tool for high-throughput investigation of RSR at the cellular level and may be useful in quality control of retinal precursor cell transplantation.
离体培养解离的视网膜神经元是研究视网膜突触再生(RSR)和探索人工视网膜潜力的重要模型。在此,将视网膜前体细胞培养在具有多个微通道阵列的微流控芯片中,以重建视网膜神经元突触。培养的视网膜细胞通过微通道进行物理连接。通过给予甘氨酸能因子证明了细胞通过微通道激活电信号转导。此外,采用基于图像的分析方法来量化突触连接并评估突触再生的动力学。当抑制剂甘氨酸浓度低于100μM时,RSR速率显著降低,然后在较高浓度下趋于相对恒定水平。此外,氯化钾化学刺激可增强RSR。总体而言,微流控突触再生芯片为在细胞水平高通量研究RSR提供了一种新型工具,可能有助于视网膜前体细胞移植的质量控制。