Multi-compartment neuron-glia co-culture platform for localized CNS axon-glia interaction study.
Multi-compartment neuron-glia co-culture platform for localized CNS axon-glia interaction study.
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DOI：
10.1039/c2lc40303j
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发表时间：
2012-09-21
期刊：
影响因子：
6.1
通讯作者：
Han A
中科院分区：
文献类型：
--
作者：
Park J;Koito H;Li J;Han A
在中枢神经系统(CNS)中，少突胶质细胞(OLs)形成髓鞘对于快速神经冲动传导是必不可少的。轴突和OL之间的相互信号协调髓鞘的发生，但在很大程度上仍然难以捉摸。在本研究中，我们提出了一种新型的多隔室CNS神经元-胶质细胞微流控共培养平台。该平台能够进行并行的局部药物和生物分子治疗，同时在单个设备中进行多种共培养条件，以更高的吞吐量研究轴突-神经胶质细胞的相互作用。这种新颖的“微-宏混合软光刻母版制造”(MMHSM)技术能够快速制造大量包含毫米级和微米级结构的精确复制的PDMS器件，而无需任何手动储存层冲压工艺。从神经元胞体生长的轴突被物理和流体隔离在六个卫星轴突/神经胶质隔室内进行局部治疗。星形胶质细胞在卫星轴突/神经胶室中分离的轴突建立后种植和共培养时，发现它们倾向于生长在轴突下面，因此对已建立的轴突层造成了物理上的破坏。相反，少突胶质前体细胞(OPC)可以与分离的轴突成功共培养，并分化为表达髓鞘碱性蛋白的成熟OPC，突起与邻近轴突一致。用高浓度神经酰胺(150μM)处理6个轴突/神经胶质室内的OPC，以确认卫星室之间的流体隔离。此外，在单个装置内用不同浓度的硫酸软骨素蛋白多糖(CSPG，0-25μg/ml)处理分离的轴突，以展示该装置平行定位的生物分子处理能力。这些结果表明，该平台可以作为研究中枢神经系统轴突生物学和轴突-神经胶质细胞相互作用的有力工具，具有局部生物分子治疗的能力。
Formation of myelin sheaths by oligodendrocytes (OLs) in the central nervous system (CNS) is essential for rapid nerve impulse conduction. Reciprocal signaling between axons and OLs orchestrates myelinogenesis but remains largely elusive. In this study, we present a novel multi-compartment CNS neuron-glia microfluidic co-culture platform. The platform is capable of conducting parallel localized drug and biomolecule treatments while carrying out multiple co-culture conditions in a single device for studying axon-glia interactions at a higher throughput. The novel “micro-macro hybrid soft-lithography master fabrication” (MMHSM) technique enables large number of precisely replicated PDMS devices incorporating both millimeter and micrometer scale structures to be rapidly fabricated without any manual reservoir punching processes. Axons grown from the neuronal somata were physically and fluidically isolated inside the six satellite axon/glia compartments for localized treatments. Astrocytes, when seeded and co-cultured after the establishment of the isolated axons in the satellite axon/glia compartments, were found to physically damage the established axonal layer as they tend to grow underneath the axons. In contrast, oligodendrocyte progenitor cells (OPCs) could be co-cultured successfully with the isolated axons and differentiated into mature myelin basic protein-expressing OLs with processes aligning to neighboring axons. OPCs inside the six axon/glia compartments were treated with high concentration of ceramide (150 μM) to confirm the fluidic isolation among satellite compartments. In addition, isolated axons were treated with varying concentrations of chondroitin sulfate proteoglycan (CSPG, 0-25 μg/ml) within a single device to demonstrate the parallel localized biomolecular treatment capability of the device. These results indicate that the proposed platform can be used as a powerful tool to study CNS axonal biology and axon-glia interactions with the capacity for localized biomolecular treatments.
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DOI：
10.1083/jcb.85.3.890
发表时间：
1980-06
期刊：
The Journal of cell biology
影响因子：
--
作者：
McCarthy KD;de Vellis J
通讯作者：
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影响因子：
6.1
作者：
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通讯作者：
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DOI：
10.1073/pnas.0502552102
发表时间：
2005-07-12
影响因子：
11.1
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通讯作者：
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影响因子：
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影响因子：
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作者：
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