Mehenti Neville Z, Tsien Greg S, Leng Theodore, Fishman Harvey A, Bent Stacey F
Department of Chemical Engineering, Stanford University, CA 94305, USA.
Biomed Microdevices. 2006 Jun;8(2):141-50. doi: 10.1007/s10544-006-7709-3.
In this work, we use cell micropatterning technologies to direct neuronal growth to individual electrodes, and demonstrate that such an approach can achieve selective stimulation and lower stimulation thresholds than current field-effect based retinal prostheses. Rat retinal ganglion cells (RGCs) were purified through immunopanning techniques, and microcontact printing (microCP) was applied to align and pattern laminin on a microelectrode array, on which the RGCs were seeded and extended neurites along the pattern to individual electrodes. The stimulation threshold currents of RGCs micropatterned to electrodes were found to be significantly less than those of non-patterned RGCs over a wide range of electrode-soma distances, as determined with calcium imaging techniques. Moreover, the stimulation threshold for micropatterned cells was found to be independent of electrode-soma distance, and there was no significant effect of microCP on cell excitability. The effects of additional stimulation parameters, such as electrode size and pulse duration, on threshold currents were determined. The stimulation results quantitatively demonstrate the potential benefits of a retinal prosthetic interface based on directed neuronal growth.
在这项工作中,我们使用细胞微图案化技术引导神经元生长至单个电极,并证明这种方法能够实现选择性刺激,且与当前基于场效应的视网膜假体相比,刺激阈值更低。通过免疫淘选技术纯化大鼠视网膜神经节细胞(RGCs),并应用微接触印刷(microCP)在微电极阵列上排列和图案化层粘连蛋白,将RGCs接种在该阵列上,使其神经突沿着图案延伸至各个电极。通过钙成像技术测定发现,在广泛的电极-胞体距离范围内,微图案化至电极的RGCs的刺激阈值电流显著低于未图案化的RGCs。此外,发现微图案化细胞的刺激阈值与电极-胞体距离无关,且微CP对细胞兴奋性无显著影响。确定了诸如电极尺寸和脉冲持续时间等额外刺激参数对阈值电流的影响。刺激结果定量地证明了基于定向神经元生长的视网膜假体接口的潜在益处。