Department of Biomedical Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Lab Chip. 2010 Mar 21;10(6):741-7. doi: 10.1039/b918640a. Epub 2010 Jan 5.
We describe a compartmentalized circular microfluidic platform that enables directed cell placement within defined microenvironments for the study of axon-glia interactions. The multi-compartment platform consists of independent units of radial microchannel arrays that fluidically isolate somal from axonal compartments. Fluidic access ports punched near the microchannels allow for direct pipetting of cells into the device. Adjacent somal or axonal compartments can be readily merged so that independent groups of neurons or axons can be maintained in either separate or uniform microenvironments. We demonstrate three distinct modes of directed cell placement in this device, to suit varying experimental needs for the study of axon-glia interactions: (1) centrifugation of the circular platform can result in a two-fold increase in axonal throughput in microchannels and provides a new technique to establish axon-glia interactions; (2) microstencils can be utilized to directly place glial cells within areas of interest; and (3) intimate axon-glia co-culture can be attained via standard pipetting techniques. We take advantage of this microfluidic platform to demonstrate a two-fold preferential accumulation of microglia specifically near injured CNS axons, an event implicated in the maintenance and progression of a number of chronic neuroinflammatory and neurodegenerative diseases.
我们描述了一种分隔的环形微流控平台,该平台能够在定义的微环境中定向放置细胞,以研究轴突-胶质相互作用。该多隔室平台由独立的径向微通道阵列单元组成,这些微通道能够将胞体与轴突隔室隔开。在微通道附近打孔的流体接入端口允许直接将细胞吸入设备中。相邻的胞体或轴突隔室可以很容易地合并,从而可以将独立的神经元或轴突群体分别或统一地保持在不同的微环境中。我们在该设备中展示了三种不同的定向细胞放置模式,以满足研究轴突-胶质相互作用的不同实验需求:(1) 环形平台的离心作用可使微通道中的轴突吞吐量增加一倍,并提供了建立轴突-胶质相互作用的新技术;(2) 可以使用微模板直接将神经胶质细胞放置在感兴趣的区域;(3) 通过标准的移液技术可以实现紧密的轴突-胶质共培养。我们利用这个微流控平台来证明小胶质细胞在中枢神经系统损伤轴突附近的两倍优先聚集,这一事件与许多慢性神经炎症和神经退行性疾病的维持和进展有关。